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\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-440\slmult0 \fs
46\cf1\f0\charscalex100 {A new approach for structure analysis}\par\pard\li1013\
ri0\sl-520\slmult0 \fs46\cf1\f0\charscalex100 {of two-dimensional membrane}\par\
pard\li1013\ri0\sl-520\slmult0 \fs46\cf1\f0\charscalex100 {protein crystals usin
g X-ray powder}\par\pard\li1013\ri0\sl-520\slmult0 \fs46\cf1\f0\charscalex100 {d
iffraction data}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri
0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard
\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf
0\par\pard\li1013\ri0\sl-386\slmult0 \fs24\cf1\f0\charscalex100 {R. A. Dilanian,
}{\super \fs16\up3\cf1\f0\charscalex100 {1}}\fs24\cf1\f0\charscalex100 {* C. Dar
manin,}{\super \fs16\up3\cf1\f0\charscalex100 {2}}\fs24\cf1\f0\charscalex100 {J.
N. Varghese,}{\super \fs16\up3\cf1\f0\charscalex100 {2}}\fs24\cf1\f0\charscalex

100 {S. W. Wilkins,}{\super \fs16\up3\cf1\f0\charscalex100 {3}}\par\pard\li1013\


ri0\sl-280\slmult0 \fs24\cf1\f0\charscalex100 {T. Oka,}{\super \fs16\up3\cf1\f0\
charscalex100 {4,5}}\fs24\cf1\f0\charscalex100 {N. Yagi,}{\super \fs16\up3\cf1\f
0\charscalex100 {5}}\fs24\cf1\f0\charscalex100 {H. M. Quiney,}{\super \fs16\up3\
cf1\f0\charscalex100 {1}}\fs24\cf1\f0\charscalex100 {and K. A. Nugent}{\super \f
s16\up3\cf1\f0\charscalex100 {1}}\par\pard\li1013\ri0\sl-373\slmult0 \fs10\cf1\f
0\charscalex100 {1}\fs16\cf1\f0\charscalex100 {ARC Centre of Excellence for Cohe
rent X-ray Science, School of Physics, The University of Melbourne, VIC 3010, Au
stralia}\par\pard\li1013\ri0\sl-240\slmult0 \fs10\cf1\f0\charscalex100 {2}\fs16\
cf1\f0\charscalex100 {ARC Centre of Excellence for Coherent X-ray Science, CSIRO
Preventative Health Flagship, CSIRO Molecular Health and}\par\pard\li1013\ri0\s
l-200\slmult0 \fs16\cf1\f0\charscalex100 {Technologies, Melbourne, VIC 3052, Aus
tralia}\par\pard\li1013\ri0\sl-240\slmult0 \fs8\cf1\f0\charscalex100 {3}\fs14\cf
1\f0\charscalex100 {ARC Centre of Excellence for Coherent X-ray Science, CSIRO M
aterials Science and Engineering, Melbourne, VIC 3169, Australia}\par\pard\li101
3\ri0\sl-240\slmult0 \fs10\cf1\f0\charscalex100 {4}\fs16\cf1\f0\charscalex100 {D
epartment of Physics, Faculty of Science and Technology, Shizuoka University, Sh
izuoka 422-8529, Japan}\par\pard\li1013\ri0\sl-240\slmult0 \fs10\cf1\f0\charscal
ex100 {5}\fs16\cf1\f0\charscalex100 {Research and Utilization Division, SPring-8
/JASRI, Hyogo 679-5198, Japan}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\
pard\li1013\ri0\sl-266\slmult0 \fs16\cf1\f0\charscalex100 {Received 27 July 2010
; Revised 10 October 2010; Accepted 24 November 2010}\par\pard\li1013\ri0\sl-200
\slmult0 \fs16\cf1\f0\charscalex100 {DOI: 10.1002/pro.572}\par\pard\li1013\ri0\s
l-200\slmult0 \fs16\cf1\f0\charscalex100 {Published online 8 December 2010 prote
inscience.org}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\
sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-360\slmult0 \fs18\cf1\f0\charsca
lex100 {Abstract: The application of powder diffraction methods to problems in s
tructural biology is}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex
100 {generally regarded as intractable because of the large number of unresolved
, overlapping X-ray}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex1
00 {reflections. Here, we use information about unit cell lattice parameters, sp
ace group}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {transf
ormations, and chemical composition as a priori information in a bootstrap proce
ss that}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {resolves
the ambiguities associated with overlapping reflections. The measured ratios of
}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {reflections tha
t can be resolved experimentally are used to refine the position, the shape, and
the}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {orientation
of low-resolution molecular structures within the unit cell, in leading to the
resolution of}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {th
e overlapping reflections. The molecular model is then made progressively more s
ophisticated as}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {
additional diffraction information is included in the analysis. We apply our met
hod to the recovery}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex1
00 {of the structure of the bacteriorhodopsin molecule (bR) to a resolution of 7
A}\fs18\cf1\f0\charscalex100 {\uc1\u730X }\fs18\cf1\f0\charscalex100 {using exp
erimental}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {data o
btained from two-dimensional purple membrane crystals. The approach can be used
to}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {determine the
structure factors directly or to provide reliable low-resolution phase informat
ion that}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {can be
refined further by the conventional methods of protein crystallography.}\par\par
d\li1013\ri0\sl-373\slmult0 \fs16\cf1\f0\charscalex100 {Keywords: integral membr
ane proteins; two-dimensional crystals; X-ray powder diffraction; pattern}\par\p
ard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {decomposition}\par\par
d\sect\sectd\sbknone\cols2\colno1\colw5786\colsr-0\colno2\colw6013\pard\li1013\r
i0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-373\slmult0 \fs18\cf1\f0\char
scalex100 {Introduction}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charsca
lex100 {Integral membrane proteins (IMPs) are involved in}\par\pard\li1013\ri0\s
l-240\slmult0 \fs16\cf1\f0\charscalex100 {such neurological processes as memory

and fine}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {motor c


ontrol. Their dysfunctions are implicated in}\par\pard\li1013\ri0\sl-240\slmult0
\fs16\cf1\f0\charscalex100 {many diseases including schizophrenia, Alzheimer\uc
1\u8217Xs}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-2
13\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex1
00 {Grant sponsors: Australian Research Council (Centres of}\par\pard\li1013\ri0
\sl-200\slmult0 \*\tx1906\*\tx2333\*\tx3213\*\tx4093\*\tx5000\fs16\cf1\f0\charsc
alex100 {Excellence}\tab \fs16\cf1\f0\charscalex100 {and}\tab \fs16\cf1\f0\chars
calex100 {Federation}\tab \fs16\cf1\f0\charscalex100 {Fellowship}\tab \fs16\cf1\
f0\charscalex100 {programs);}\tab \fs16\cf1\f0\charscalex100 {CSIRO}\par\pard\li
1013\ri0\sl-200\slmult0 \*\tx2013\*\tx2613\*\tx3386\*\tx4853\fs16\cf1\f0\charsca
lex100 {Preventative}\tab \fs16\cf1\f0\charscalex100 {Health}\tab \fs16\cf1\f0\c
harscalex100 {Flagship;}\tab \fs16\cf1\f0\charscalex100 {Neurodegenerative}\tab
\fs16\cf1\f0\charscalex100 {Diseases}\par\pard\li1013\ri0\sl-200\slmult0 \fs16\c
f1\f0\charscalex100 {Theme; CSIRO; Emerging Science Initiative.}\par\pard\li1013
\ri0\sl-306\slmult0 \*\tx2586\fs16\cf1\f0\charscalex100 {*}\fs16\cf1\f0\charscal
ex100 {Correspondence to:}\tab \fs16\cf1\f0\charscalex100 {R. A. Dilanian, Schoo
l of Physics, The}\par\pard\li1013\ri0\sl-173\slmult0 \fs16\cf1\f0\charscalex100
{University of Melbourne, VIC 3010, Australia.}\par\pard\li1013\ri0\sl-200\slmu
lt0 \fs16\cf1\f0\charscalex100 {E-mail: roubend@unimelb.edu.au}\par\pard\li1013\
ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pa
rd\li1013\ri0\sl-266\slmult0 \*\tx2946\fs12\cf1\f0\charscalex100 {Published by W
iley-Blackwell.}{ }\fs12\cf1\f0\charscalex100 {V}\tab \fs12\cf1\f0\charscalex100
{2010 The Protein Society}\par\column\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par
\pard\li0\ri0\sl-360\slmult0 \*\tx3173\*\tx4120\fs18\cf1\f0\charscalex100 {disea
se, and Parkinson\uc1\u8217Xs disease.}\fs12\cf1\f0\charscalex100 {1\uc1\u8211X5
}\tab \fs18\cf1\f0\charscalex100 {In 2005,}\tab \fs18\cf1\f0\charscalex100 {60%}
\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {of the available ma
rketed pharmaceuticals targeted}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\ch
arscalex100 {membrane proteins,}{\super \fs12\up3\cf1\f0\charscalex100 {6}}\fs18
\cf1\f0\charscalex100 {a proportion that one would}\par\pard\li0\ri0\sl-240\slmu
lt0 \fs18\cf1\f0\charscalex100 {expect to increase with further research of IMP}
\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {classes and subclas
ses. Different isoforms exist for}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\
charscalex100 {many of the IMPs, and the design of new drugs can}\par\pard\li0\r
i0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {only be based on known structures
and information}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {abo
ut the binding mechanisms. Despite this impor-}\par\pard\li0\ri0\sl-240\slmult0
\*\tx1240\fs18\cf1\f0\charscalex100 {tance, only}\tab \fs18\cf1\f0\charscalex100
{30 unique structures of IMPs have}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f
0\charscalex100 {been solved to atomic resolution, compared with}\par\pard\li0\r
i0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {more than 3000 unique crystal stru
ctures for soluble}\par\pard\li0\ri0\sl-240\slmult0 \*\tx906\*\tx1733\*\tx2053\*
\tx2506\*\tx3373\*\tx4306\fs18\cf1\f0\charscalex100 {proteins,}\tab \fs18\cf1\f0
\charscalex100 {because}\tab \fs18\cf1\f0\charscalex100 {of}\tab \fs18\cf1\f0\ch
arscalex100 {the}\tab \fs18\cf1\f0\charscalex100 {intrinsic}\tab \fs18\cf1\f0\ch
arscalex100 {difficulty}\tab \fs18\cf1\f0\charscalex100 {in}\par\pard\li0\ri0\sl
-240\slmult0 \fs18\cf1\f0\charscalex100 {extracting these proteins from the memb
rane and}\par\pard\li1506\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1506\ri0\sl-21
3\slmult0 \fs18\cf0\par\pard\li1506\ri0\sl-293\slmult0 \*\tx4253\fs12\cf1\f0\cha
rscalex100 {PROTEIN SCIENCE 2011 VOL 20:457\uc1\u8212X464}\tab \fs18\cf1\f0\char
scalex100 {457}\par\pard\sect\sectd\sbknone\pard\sect\sectd\sbkpage\pgwsxn12600\
pghsxn16200\marglsxn666\margrsxn133\margtsxn666\margbsxn173\cols2\colno1\colw578
6\colsr-0\colno2\colw6013\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li10
13\ri0\sl-200\slmult0 \fs18\cf1\f0\charscalex100 {manipulating them to produce t
hree-dimensional}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100
{(3D) crystals for structural analysis. The majority of}\par\pard\li1013\ri0\sl240\slmult0 \fs16\cf1\f0\charscalex100 {known IMPs are much more likely to form
two-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {dimensional
(2D), rather than 3D crystals during}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\

cf1\f0\charscalex100 {the crystallization processes,}{\super \fs10\up3\cf1\f0\ch


arscalex100 {7}}\fs16\cf1\f0\charscalex100 {which limits the possi-}\par\pard\li
1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {bility of obtaining their mo
lecular structures using}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charsc
alex100 {the standard methods of protein crystallography.}\par\pard\li1013\ri0\s
l-240\slmult0 \fs16\cf1\f0\charscalex100 {Powder diffraction methods are, in con
trast, not crit-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100
{ically sensitive to the quality and dimensions of}\par\pard\li1013\ri0\sl-240\s
lmult0 \fs16\cf1\f0\charscalex100 {crystals, and this suggests their use in the
structure}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {analys
is of 2D crystals.}\par\pard\li1373\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex10
0 {During the last decade, protein structures have}\par\pard\li1013\ri0\sl-240\s
lmult0 \fs18\cf1\f0\charscalex100 {been determined by X-ray diffraction from sub
mi-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {cron protein
crystals that form a powder-like sample}\par\pard\li1013\ri0\sl-240\slmult0 \fs
18\cf1\f0\charscalex100 {(see review articles}{\super \fs12\up3\cf1\f0\charscale
x100 {8,9}}\fs18\cf1\f0\charscalex100 {), an idea originally demon-}\par\pard\li
1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {strated to be feasible in 19
99, when the first protein}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\char
scalex100 {structure refinement was reported.}{\super \fs12\up3\cf1\f0\charscale
x100 {10}}\fs18\cf1\f0\charscalex100 {According to}\par\pard\li1013\ri0\sl-240\s
lmult0 \fs18\cf1\f0\charscalex100 {more recent studies, the synchrotron X-ray po
wder}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {diffraction
technique has been successfully adapted}\par\pard\li1013\ri0\sl-240\slmult0 \fs
16\cf1\f0\charscalex100 {to various proteins, using state of the art instrumen-}
\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {tation and algor
ithms. Recent progress in powder}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f
0\charscalex100 {diffraction data collection}{\super \fs10\up3\cf1\f0\charscalex
100 {11}}\fs16\cf1\f0\charscalex100 {and interpretation}{\super \fs10\up3\cf1\f0
\charscalex100 {9}}\fs16\cf1\f0\charscalex100 {has}\par\pard\li1013\ri0\sl-240\s
lmult0 \fs16\cf1\f0\charscalex100 {facilitated the study of several biologically
interest-}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx2520\fs16\cf1\f0\charscalex1
00 {ing proteins.}\fs10\cf1\f0\charscalex100 {12\uc1\u8211X18}\tab \fs16\cf1\f0\
charscalex100 {The successful solution of different}\par\pard\li1013\ri0\sl-240\
slmult0 \fs18\cf1\f0\charscalex100 {protein structures has been achieved via tra
ditional}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {methods
in single-crystal crystallography, such as}\par\pard\li1013\ri0\sl-240\slmult0
\*\tx4240\fs16\cf1\f0\charscalex100 {molecular replacement (MR),}\fs10\cf1\f0\ch
arscalex100 {11,13,17}\tab \fs16\cf1\f0\charscalex100 {single isomor-}\par\pard\
li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {phous replacement revealin
g the molecular enve-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscale
x100 {lope,}{\super \fs10\up3\cf1\f0\charscalex100 {14}}\fs16\cf1\f0\charscalex1
00 {and multiple isomorphous replacement show-}\par\pard\li1013\ri0\sl-240\slmul
t0 \*\tx1453\*\tx2480\*\tx3426\*\tx4360\*\tx4693\*\tx4973\fs18\cf1\f0\charscalex
100 {ing}\tab \fs18\cf1\f0\charscalex100 {secondary}\tab \fs18\cf1\f0\charscalex
100 {structure}\tab \fs18\cf1\f0\charscalex100 {elements}\tab \fs18\cf1\f0\chars
calex100 {of}\tab \fs18\cf1\f0\charscalex100 {a}\tab \fs18\cf1\f0\charscalex100
{model}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {protein.}
{\super \fs12\up3\cf1\f0\charscalex100 {15}}\par\pard\li1373\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {The inevitable loss of information arising from}\par
\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {the collapse of the
3D reciprocal space onto a one-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0
\charscalex100 {dimensional powder diffraction pattern leads, how-}\par\pard\li1
013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {ever, to overlapping reflecti
ons (partial or complete)}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\chars
calex100 {in the data, which is an obvious disadvantage with}\par\pard\li1013\ri
0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {respect to single crystal diffracti
on. This is the main}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex
100 {reason why the methods of powder diffraction crys-}\par\pard\li1013\ri0\sl240\slmult0 \fs18\cf1\f0\charscalex100 {tallography are still not as popular for

the structure}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {a


nalysis of proteins as single crystal crystallography.}\par\pard\li1013\ri0\sl-2
40\slmult0 \fs18\cf1\f0\charscalex100 {Rapid growth in demand for structural inf
ormation}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {of memb
rane proteins will certainly lead to a more}\par\pard\li1013\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {intensive use of approaches developed for powder}\pa
r\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {diffraction crystal
lography if they can be made to}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0
\charscalex100 {work reliably.}\par\pard\li1373\ri0\sl-240\slmult0 \fs16\cf1\f0\
charscalex100 {The complete overlapping of reflections can be}\par\pard\li1013\r
i0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {either accidental, which is common
for crystals with}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex10
0 {low lattice symmetry as a result of the peak broad-}\par\pard\li1013\ri0\sl-2
40\slmult0 \fs18\cf1\f0\charscalex100 {ening effect and/or limitation in instrum
ental resolu-}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {ti
on, or exact in the case of high lattice symmetry.}\par\pard\li1013\ri0\sl-240\s
lmult0 \fs16\cf1\f0\charscalex100 {Preliminary analysis of available structures
of 2D}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {protein cr
ystals shows that they crystallize preferen-}\par\pard\li1013\ri0\sl-240\slmult0
\*\tx4640\*\tx4933\*\tx5186\fs16\cf1\f0\charscalex100 {tially with low lattice
symmetry, such as}\tab \fs16\cf1\f0\charscalex100 {p2}\tab \fs16\cf1\f0\charscal
ex100 {or}\tab \fs16\cf1\f0\charscalex100 {cm}\fs16\cf1\f0\charscalex100 {,}\par
\pard\li1013\ri0\sl-240\slmult0 \*\tx3920\*\tx4480\fs16\cf1\f0\charscalex100 {or
with high symmetry, such as}\tab \fs16\cf1\f0\charscalex100 {p}\fs16\cf1\f0\cha
rscalex100 {3 or}\tab \fs16\cf1\f0\charscalex100 {p}\fs16\cf1\f0\charscalex100 {
6 (see, for}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx3026\fs16\cf1\f0\charscalex
100 {example, articles}\fs10\cf1\f0\charscalex100 {19\uc1\u8211X22}\tab \fs16\cf
1\f0\charscalex100 {and references therein). It}\par\pard\li1013\ri0\sl-240\slmu
lt0 \fs18\cf1\f0\charscalex100 {should be noted that for plane symmetries, such
as}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx1373\*\tx2160\fs16\cf1\f0\charscalex
100 {p}\fs16\cf1\f0\charscalex100 {4,}\tab \fs16\cf1\f0\charscalex100 {p}\fs16\c
f1\f0\charscalex100 {3, and}\tab \fs16\cf1\f0\charscalex100 {p}\fs16\cf1\f0\char
scalex100 {6, all (}\fs16\cf1\f0\charscalex100 {h}\fs16\cf1\f0\charscalex100 {,}
\fs16\cf1\f0\charscalex100 {k}\fs16\cf1\f0\charscalex100 {) and (}\fs16\cf1\f0\c
harscalex100 {k}\fs16\cf1\f0\charscalex100 {,}\fs16\cf1\f0\charscalex100 {h}\fs1
6\cf1\f0\charscalex100 {) reflections are}\par\pard\li1013\ri0\sl-240\slmult0 \*
\tx3613\fs18\cf1\f0\charscalex100 {exactly overlapped and form}\tab \fs18\cf1\f0
\charscalex100 {50% of the total num-}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\
cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-293\slmu
lt0 \*\tx1506\fs18\cf1\f0\charscalex100 {458}\tab \fs12\cf1\f0\charscalex100 {PR
OTEINSCIENCE.ORG}\par\column\pard\li0\ri0\sl-413\slmult0 \fs18\cf1\f0\charscalex
100 {ber of diffraction peaks up to 7-A}\fs18\cf1\f0\charscalex100 {\uc1\u730X }
\fs18\cf1\f0\charscalex100 {resolution. This}\par\pard\li0\ri0\sl-240\slmult0 \f
s18\cf1\f0\charscalex100 {statistic would appear to render structure analysis}\p
ar\pard\li0\ri0\sl-240\slmult0 \*\tx613\*\tx1453\*\tx2533\*\tx3133\*\tx3813\fs18
\cf1\f0\charscalex100 {from}\tab \fs18\cf1\f0\charscalex100 {powder}\tab \fs18\c
f1\f0\charscalex100 {diffraction}\tab \fs18\cf1\f0\charscalex100 {data}\tab \fs1
8\cf1\f0\charscalex100 {using}\tab \fs18\cf1\f0\charscalex100 {existing}\par\par
d\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {approaches practically impo
ssible if no prior struc-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscal
ex100 {ture information exists.}\par\pard\li373\ri0\sl-240\slmult0 \fs18\cf1\f0\
charscalex100 {Various methods have been developed to over-}\par\pard\li0\ri0\sl
-240\slmult0 \*\tx3973\fs18\cf1\f0\charscalex100 {come the overlapping reflectio
n problem.}\fs12\cf1\f0\charscalex100 {23\uc1\u8211X29}\tab \fs18\cf1\f0\charsca
lex100 {These}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {inclu
de, for example, the squaring method,}{\super \fs12\up3\cf1\f0\charscalex100 {23
}}\fs18\cf1\f0\charscalex100 {the fast}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf
1\f0\charscalex100 {iterative Patterson squaring method,}{\super \fs12\up3\cf1\f
0\charscalex100 {26}}\fs18\cf1\f0\charscalex100 {or the maxi-}\par\pard\li0\ri0\
sl-240\slmult0 \fs18\cf1\f0\charscalex100 {mum entropy Patterson (MEP) method.}{

\super \fs12\up3\cf1\f0\charscalex100 {24}}\fs18\cf1\f0\charscalex100 {Although}


\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {there are many exam
ples of successful decomposi-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\char
scalex100 {tion using these approaches, all of them frequently}\par\pard\li0\ri0
\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {fail to separate individual componen
ts correctly}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {when t
he number of overlapped reflection becomes}\par\pard\li0\ri0\sl-240\slmult0 \fs1
8\cf1\f0\charscalex100 {comparable with the number of well-resolved reflec-}\par
\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {tions. Integrated inten
sities of overlapped reflections}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\c
harscalex100 {in powder diffraction data can be further improved}\par\pard\li0\r
i0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {by the iterative combination of th
e Le Bail analysis}{\super \fs12\up3\cf1\f0\charscalex100 {30}}\par\pard\li0\ri0
\sl-240\slmult0 \*\tx3333\fs18\cf1\f0\charscalex100 {and the MEP-based approache
s.}\fs12\cf1\f0\charscalex100 {24,31}\tab \fs18\cf1\f0\charscalex100 {Such a com
-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {bined approach to
analysis is particularly effective}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f
0\charscalex100 {for deriving correct intensity ratios of the individual}\par\pa
rd\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {components of the overlapp
ing reflections for com-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscale
x100 {pounds containing relatively heavy atoms. A differ-}\par\pard\li0\ri0\sl-2
40\slmult0 \fs18\cf1\f0\charscalex100 {ent approach, involving the combination o
f the Le}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {Bail analy
sis and the charge-flipping approach, also}\par\pard\li0\ri0\sl-240\slmult0 \fs1
8\cf1\f0\charscalex100 {allows the intensities of overlapping reflections to be}
\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {resolved.}{\super \
fs12\up3\cf1\f0\charscalex100 {32}}\fs18\cf1\f0\charscalex100 {The approach deal
s efficiently with the}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex1
00 {overlapping problem and provides a fast and effec-}\par\pard\li0\ri0\sl-240\
slmult0 \*\tx493\*\tx1013\*\tx1426\*\tx2266\*\tx3053\*\tx4093\fs18\cf1\f0\charsc
alex100 {tive}\tab \fs18\cf1\f0\charscalex100 {way}\tab \fs18\cf1\f0\charscalex1
00 {for}\tab \fs18\cf1\f0\charscalex100 {phasing}\tab \fs18\cf1\f0\charscalex100
{powder}\tab \fs18\cf1\f0\charscalex100 {diffraction}\tab \fs18\cf1\f0\charscal
ex100 {data}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {obtaine
d from inorganic materials with relatively}\par\pard\li0\ri0\sl-240\slmult0 \fs1
8\cf1\f0\charscalex100 {small unit cell dimensions. It has not yet been estab-}\
par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {lished, however, tha
t a similar approach is effective}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\
charscalex100 {in cases in which the unit cells are large, such as}\par\pard\li0
\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {with protein crystals.}\par\pard
\li373\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {In this article, we demons
trate a new iterative}\par\pard\li0\ri0\sl-240\slmult0 \*\tx973\*\tx1333\*\tx232
0\*\tx2960\*\tx4093\fs18\cf1\f0\charscalex100 {approach}\tab \fs18\cf1\f0\charsc
alex100 {to}\tab \fs18\cf1\f0\charscalex100 {overcome}\tab \fs18\cf1\f0\charscal
ex100 {these}\tab \fs18\cf1\f0\charscalex100 {difficulties.}\tab \fs18\cf1\f0\ch
arscalex100 {This}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {a
pproach allows severely or completely overlapping}\par\pard\li0\ri0\sl-240\slmul
t0 \fs18\cf1\f0\charscalex100 {reflections to be decomposed using intensity rati
os of}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {resolved refl
ections and a priori information, such as}\par\pard\li0\ri0\sl-240\slmult0 \fs18
\cf1\f0\charscalex100 {unit cell parameters, information about the crystal-}\par
\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {lographic symmetry grou
p, and the chemical compo-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charsca
lex100 {sition of the sample. This approach not only allows}\par\pard\li0\ri0\sl
-240\slmult0 \fs18\cf1\f0\charscalex100 {the intensity of the individual compone
nts of the dif-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {fra
ction pattern to be extracted but also furnishes}\par\pard\li0\ri0\sl-240\slmult
0 \fs18\cf1\f0\charscalex100 {initial estimates of the phases for subsequent}\pa
r\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {refinement.}\par\pard\
li373\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {The bacteriorhodopsin molec

ule (bR), whose}\par\pard\li0\ri0\sl-240\slmult0 \*\tx3880\fs18\cf1\f0\charscale


x100 {structure has previously been analyzed,}\fs12\cf1\f0\charscalex100 {22,33}
\tab \fs18\cf1\f0\charscalex100 {will be}\par\pard\li0\ri0\sl-240\slmult0 \fs18\
cf1\f0\charscalex100 {used as the target system of our analysis. The crys-}\par\
pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {talline form of a bR mol
ecule, the 2D crystal of pur-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\char
scalex100 {ple membrane (PM), has a plane group symmetry}\par\pard\li0\ri0\sl-24
0\slmult0 \fs18\cf1\f0\charscalex100 {p3}\fs18\cf1\f0\charscalex100 {. In this a
rticle, we demonstrate the ability of the}\par\pard\li0\ri0\sl-240\slmult0 \fs18
\cf1\f0\charscalex100 {approach to extract the individual components of}\par\par
d\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {pairs of exactly overlapped
reflections, (}\fs18\cf1\f0\charscalex100 {hk}\fs18\cf1\f0\charscalex100 {) and
(}\fs18\cf1\f0\charscalex100 {kh}\fs18\cf1\f0\charscalex100 {).}\par\pard\li0\r
i0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {The proposed approach, however, is
also applicable}\par\pard\li0\ri0\sl-240\slmult0 \*\tx360\*\tx1453\*\tx2333\*\t
x3440\fs18\cf1\f0\charscalex100 {to}\tab \fs18\cf1\f0\charscalex100 {decompose}\
tab \fs18\cf1\f0\charscalex100 {severely}\tab \fs18\cf1\f0\charscalex100 {overla
pped}\tab \fs18\cf1\f0\charscalex100 {(unresolved)}\par\pard\li0\ri0\sl-240\slmu
lt0 \fs18\cf1\f0\charscalex100 {reflections caused by low lattice symmetry or re
solu-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {tion limits.}
\par\pard\li533\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li533\ri0\sl-213\slmult0 \
fs18\cf0\par\pard\li533\ri0\sl-266\slmult0 \fs12\cf1\f0\charscalex100 {A New App
roach for Structure Analysis of 2D Membrane Proteins}\par\pard\sect\sectd\sbkpag
e
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095AC3AAF89ECF58D0ECB51D334FB30EB77F6CBD321B82CA42A840EC3863924FE1401D27C4CD0E5D
6BC117FF0062DC9A85AC7F68B778D8AB65792B91CF2A08C572DFF0903F8A3C2F73E26D360DA349D1

18C2123F945DB26582819CF961463D0B57AD380CA4300411C83E9DEB2F46F0FE97A169274CD3AD23
8ACF73B18BA8258FCD9CFE5F4A00F3AD3B4FD06E747F0E7882D355B7B7B98DE3647B555FB45DCAC1
434533025A4249F9BF3ACDF0DDBF877C63F0FE6D6B5FD49ED75A86EDEE6F2FD2545BAB7646384424
12A9B780B5DFE93F0D7C2DA2EB0BAA58E9DB26472F12172D1C4C4609443C29C7715149F0B3C232EB
2FAA3696BE649209248558885D87731FDD3CF38C75A00E5B4B834CB4F893E35BFB6D3ACFCCB7B2B5
BAB40F0282256898E40C643312338E49AC88B4CD3355F81AFE274988D7A28A4BB7D54BE2E04EAE72
A645E71FC2173D315EA73F83744B8F128D7E5B526FD5154B6E3B1F6FDD2CBD091C633D302A92FC39
F0E2CF2B476D3C76F34BE74B6293B2DB3BE7A9881DA79038C50070D0E8F6FE23F891A22EB56ACCD7
BE1813DEC24B209242C01DC33EFF009D7A75DDAAE9FE17B9B4B08DA3482CDD2058F395C21DA077CD
46DE14D21BC449AE9865FED251B56513B8C2FF007719C6DF6E95B7B47AF1401E71F055EDA1F85161
224AC143CCD3B39E11839CFE18AE7357F0DEB7E35F04EB7A90B3B66FED497EDD68CF2B899638C110
A85DA472A09C67FE5A575BA2F839B496D53C3934665F0EDF4CF756DE54C51E0CE3CC89F047C9B88D
BB7B677575567A158E9FA28D2AD7CF8AD150AA013BEE45F456CE47E140185F0CFC4BFF0009578174
EBD925DF75127D9EE8771227073F5186FC6BB2AE7B40F07687E189656D1ED9ED7CC1FBC8D656287D
CA938CFBF5AE86803CC3E3B6D1F0E0B02E265BD84C1B7A97C9FE99AF48B5DC6D212F9DFB06ECF5CE
39AE4FC45A05DF8AFC47A6DADD4689A1E9B3ADDCCACDF35CCA01D8BB790507F16EE4F6AEC85002D1
45140051451400514514005145140051451400514514005145140051451400514514005145140051
451401FFD9}
}}}}\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\margtsxn666\margbsxn173\par
d\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\c
f1\f0\charscalex100 {Results and Discussion}\par\pard\li1013\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {We here consider a cluster of atoms in the unit cell
}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {as the constitu
ent element of the crystal structure.}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\
cf1\f0\charscalex100 {The cluster can include several, single, or even part}\par
\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {of the membrane prot
ein. It contains no internal}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\ch
arscalex100 {symmetry operation except the identity operator.}\par\pard\li1013\r
i0\sl-240\slmult0 \*\tx1493\*\tx2733\*\tx3146\*\tx3640\*\tx4120\*\tx4613\*\tx537
3\fs18\cf1\f0\charscalex100 {For}\tab \fs18\cf1\f0\charscalex100 {convenience,}\
tab \fs18\cf1\f0\charscalex100 {we}\tab \fs18\cf1\f0\charscalex100 {will}\tab \f
s18\cf1\f0\charscalex100 {call}\tab \fs18\cf1\f0\charscalex100 {this}\tab \fs18\
cf1\f0\charscalex100 {cluster}\tab \fs18\cf1\f0\charscalex100 {a}\par\pard\li101
3\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {\uc1\u8216X\uc1\u8216Xmolecule.
\uc1\u8217X\uc1\u8217X}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li
1013\ri0\sl-293\slmult0 \fs18\cf1\f0\charscalex100 {Molecular form factor and th
e resolution limit}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex10
0 {The relation between the intensity of a diffraction}\par\pard\li1013\ri0\sl-2
40\slmult0 \fs18\cf1\f0\charscalex100 {peak and the crystal structure is}\par\pa
rd\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols2\coln
o1\colw5253\colsr-0\colno2\colw6546\pard\li2200\ri0\sl-213\slmult0 \fs18\cf0\par
\pard\li2200\ri0\sl-320\slmult0 \*\tx2786\*\tx3173\*\tx4080\fs18\cf1\f0\charscal
ex100 {I}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charscalex100 {S}\tab \fs18\cf
1\f0\charscalex100 {K}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charscalex100 {S}
\tab \fs18\cf1\f0\charscalex100 {L}{\sub \fs12\cf1\f0\charscalex100 {P}}\fs18\cf
1\f0\charscalex100 { }\fs18\cf1\f0\charscalex100 {S}{ }\fs18\cf1\f0\charscalex100
{j}\fs18\cf1\f0\charscalex100 {F}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charsc
alex100 {S}\tab \fs18\cf1\f0\charscalex100 {j}\fs12\cf1\f0\charscalex100 {2}{\su
b \fs18\cf1\f0\charscalex100 {;}}\par\column\pard\li0\ri0\sl-213\slmult0 \fs18\c
f0\par\pard\li0\ri0\sl-293\slmult0 \fs18\cf1\f0\charscalex100 {(1)}\par\pard\sec
t\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\pard\li1013\ri0\
sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-253\slmult0 \fs18\cf1\f0\charsca
lex100 {where}{ }\fs18\cf1\f0\charscalex100 {S}{ }\fs18\cf1\f0\charscalex100 {is
the scattering vector,}{ }\fs18\cf1\f0\charscalex100 {K}\fs18\cf1\f0\charscalex
100 {(}\fs18\cf1\f0\charscalex100 {S}\fs18\cf1\f0\charscalex100 {) is the Laue i
n-}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx2760\fs16\cf1\f0\charscalex100 {terf
erence function,}\tab \fs16\cf1\f0\charscalex100 {L}\fs10\cf1\f0\charscalex100 {
P}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscalex100 {S}\fs16\cf1\f0\chars
calex100 {) is the Lorentz and polar-}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx2
626\fs16\cf1\f0\charscalex100 {ization factor, and}\tab \fs16\cf1\f0\charscalex1

00 {F}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscalex100 {S}\fs16\cf1\f0\c


harscalex100 {) is the structure factor. If we}\par\pard\li1013\ri0\sl-213\slmul
t0 \fs16\cf1\f0\charscalex100 {consider an individual molecule as a constituent
ele-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {ment of the
crystal structure and assume that the}\par\pard\li1013\ri0\sl-240\slmult0 \fs18
\cf1\f0\charscalex100 {asymmetric unit contains only one molecule then}\par\pard
\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {the structure factor can
be written as,}\par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\
margrsxn133\cols5\colno1\colw2346\colsr-0\colno2\colw280\colsr-0\colno3\colw1946
\colsr-0\colno4\colw680\colsr-0\colno5\colw6546\pard\li1720\ri0\sl-213\slmult0 \
fs18\cf0\par\pard\li1720\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1720\ri0\sl-240
\slmult0 \fs18\cf1\f0\charscalex100 {F}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\
charscalex100 {S}\par\column\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\
ri0\sl-213\slmult0 \fs18\cf1\f0\charscalex100 {X}\fs12\cf1\f0\charscalex100 {M}\
par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-186\slmult0 \fs12\
cf1\f0\charscalex100 {k}\fs12\cf1\f0\charscalex100 {}\fs12\cf1\f0\charscalex100 {
1}\par\column\pard\li786\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li786\ri0\sl-160\
slmult0 \fs18\cf1\f0\charscalex100 {n}\par\pard\li0\ri0\sl-293\slmult0 \*\tx506\
*\tx906\*\tx1280\fs18\cf1\f0\charscalex100 {U}{\sub \fs12\cf1\f0\charscalex100 {
k}}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charscalex100 {S}\tab \fs18\cf1\f0\c
harscalex100 {exp}\tab \fs18\cf1\f0\charscalex100 {i}\fs18\cf1\f0\charscalex100
{2}\fs18\cf1\f0\charscalex100 {p}\tab \fs18\cf1\f0\charscalex100 {S}\fs18\cf1\f0
\charscalex100 {G}\fs18\cf1\f0\charscalex100 {^}{\sub \fs12\cf1\f0\charscalex100
{k}}\fs18\cf1\f0\charscalex100 {R}{\sub \fs12\cf1\f0\charscalex100 {0}}\par\col
umn\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-253\slmult0 \fs18\
cf1\f0\charscalex100 {o}\par\pard\li120\ri0\sl-240\slmult0 \fs18\cf1\f0\charscal
ex100 {;}\par\column\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-2
13\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf1\f0\charscalex100
{(2)}\par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn13
3\cols2\colno1\colw5786\colsr-0\colno2\colw6013\pard\li1013\ri0\sl-213\slmult0 \
fs18\cf0\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {where}{
}\fs18\cf1\f0\charscalex100 {M}{ }\fs18\cf1\f0\charscalex100 {is the number of e
lements of the symmetry}\par\pard\li1013\ri0\sl-293\slmult0 \*\tx1653\*\tx5200\f
s18\cf1\f0\charscalex100 {group,}\tab \fs18\cf1\f0\charscalex100 {G}\fs18\cf1\f0
\charscalex100 {^}{\sub \fs12\cf1\f0\charscalex100 {k}}\fs18\cf1\f0\charscalex10
0 {is the matrix representation of the}\tab \fs18\cf1\f0\charscalex100 {k}\fs18\
cf1\f0\charscalex100 {th}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charsc
alex100 {symmetry element of the symmetry group, and}{ }\fs18\cf1\f0\charscalex1
00 {R}{\sub \fs12\cf1\f0\charscalex100 {0}}\fs18\cf1\f0\charscalex100 {is}\par\p
ard\li1013\ri0\sl-186\slmult0 \fs18\cf1\f0\charscalex100 {the origin of the mole
cule in the asymmetric unit.}\par\pard\li1013\ri0\sl-266\slmult0 \fs18\cf1\f0\ch
arscalex100 {U}\fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {S}\fs18
\cf1\f0\charscalex100 {) defines the scattering from the individual mole-}\par\p
ard\li1013\ri0\sl-213\slmult0 \fs18\cf1\f0\charscalex100 {cule or the molecular
form factor (MFF),}\par\pard\li2333\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li2333
\ri0\sl-266\slmult0 \*\tx3413\*\tx4653\fs18\cf1\f0\charscalex100 {X}\fs12\cf1\f0
\charscalex100 {N}\tab \fs18\cf1\f0\charscalex100 {n}\tab \fs18\cf1\f0\charscale
x100 {o}\par\pard\li1626\ri0\sl-293\slmult0 \*\tx2626\*\tx3533\*\tx3893\*\tx5253
\fs18\cf1\f0\charscalex100 {U}{\sub \fs12\cf1\f0\charscalex100 {k}}\fs18\cf1\f0\
charscalex100 { }\fs18\cf1\f0\charscalex100 {S}\tab \fs18\cf1\f0\charscalex100 {f}
{\sub \fs12\cf1\f0\charscalex100 {j}}\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0
\charscalex100 {S}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {
exp}\tab \fs18\cf1\f0\charscalex100 {i}\fs18\cf1\f0\charscalex100 {2}\fs18\cf1\f
0\charscalex100 {p}\tab \fs18\cf1\f0\charscalex100 {S}\fs18\cf1\f0\charscalex100
{G}\fs18\cf1\f0\charscalex100 {^}{\sub \fs12\cf1\f0\charscalex100 {k}}\fs18\cf1
\f0\charscalex100 {D}\fs18\cf1\f0\charscalex100 {R}{\sub \fs12\cf1\f0\charscalex
100 {j}}\fs18\cf1\f0\charscalex100 {;}\tab \fs18\cf1\f0\charscalex100 {(3)}\par\
pard\li2360\ri0\sl-146\slmult0 \fs12\cf1\f0\charscalex100 {j}\fs12\cf1\f0\charsc
alex100 {}\fs12\cf1\f0\charscalex100 {1}\par\pard\li1013\ri0\sl-386\slmult0 \*\tx
1600\*\tx1826\fs16\cf1\f0\charscalex100 {where}\tab \fs16\cf1\f0\charscalex100 {

N}\tab \fs16\cf1\f0\charscalex100 {is the number of atoms in the asymmetric}\par


\pard\li1013\ri0\sl-253\slmult0 \*\tx1493\*\tx5240\fs18\cf1\f0\charscalex100 {un
it,}\tab \fs18\cf1\f0\charscalex100 {f}{\sub \fs12\cf1\f0\charscalex100 {j}}\fs1
8\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {S}\fs18\cf1\f0\charscalex1
00 {) is the atomic scattering factor of the}\tab \fs18\cf1\f0\charscalex100 {j}
\fs18\cf1\f0\charscalex100 {th}\par\pard\li1013\ri0\sl-266\slmult0 \*\tx1560\*\t
x2173\fs18\cf1\f0\charscalex100 {atom,}\tab \fs18\cf1\f0\charscalex100 {D}\fs18\
cf1\f0\charscalex100 {R}{\sub \fs12\cf1\f0\charscalex100 {j}}\fs18\cf1\f0\charsc
alex100 {}\tab \fs18\cf1\f0\charscalex100 {R}{\sub \fs12\cf1\f0\charscalex100 {j}
}\fs18\cf1\f0\charscalex100 {R}{\sub \fs12\cf1\f0\charscalex100 {0}}\fs18\cf1\f0
\charscalex100 {is the relative position of the}\par\pard\li1013\ri0\sl-200\slmu
lt0 \fs18\cf1\f0\charscalex100 {j}\fs18\cf1\f0\charscalex100 {th atom in the asy
mmetric unit.}\par\pard\li1373\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {In
this representation, the diffraction pattern of}\par\pard\li1013\ri0\sl-240\slm
ult0 \fs16\cf1\f0\charscalex100 {the crystal can be regarded as the superpositio
n of the}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {scatter
ing from the individual molecules, the MFF and}\par\pard\li1013\ri0\sl-240\slmul
t0 \fs18\cf1\f0\charscalex100 {the Laue interference function. It has been shown
}{\super \fs12\up3\cf1\f0\charscalex100 {34}}\par\pard\li1013\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {that the powder diffraction pattern exhibits a sens
itiv-}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {ity to the
one-electron density of the molecule. In the}\par\pard\li1013\ri0\sl-240\slmult
0 \fs18\cf1\f0\charscalex100 {case of the low-resolution imaging of a scattering
}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {object of chara
cteristic dimension,}{ }\fs18\cf1\f0\charscalex100 {D}\fs18\cf1\f0\charscalex100
{, scattering from a}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscale
x100 {molecule comprised of discrete atoms is almost indis-}\par\pard\li1013\ri0
\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {tinguishable from that of an homogen
eous and contin-}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100
{uous envelope distribution of electron density if the}\par\pard\li1013\ri0\sl-2
80\slmult0 \*\tx2240\fs18\cf1\f0\charscalex100 {criterion SD}\tab \fs18\cf1\f0\c
harscalex100 {1 is satisfied, where}{ }\fs18\cf1\f0\charscalex100 {S}{ }\fs18\cf
1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {|}\fs18\cf1\f0\charscalex100
{S}\fs18\cf1\f0\charscalex100 {|.}\par\pard\li1373\ri0\sl-200\slmult0 \fs18\cf1\
f0\charscalex100 {The low-resolution shape of the molecule can be}\par\pard\li10
13\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {obtained by looking at the for
m of the asymmetric}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex1
00 {unit. Assuming that the size of the bR molecule is}\par\pard\li1013\ri0\sl-2
40\slmult0 \fs16\cf1\f0\charscalex100 {comparable with the size of the asymmetri
c unit, we}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {can c
onsider the continuous representation of the}\par\pard\li1013\ri0\sl-213\slmult0
\fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-2
66\slmult0 \fs12\cf1\f0\charscalex100 {Dilanian et al.}\par\column\pard\li0\ri0\
sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\
ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\
li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\p
ard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-186\slmult0 \*\tx760\fs
16\cf1\f0\charscalex100 {Figure 1.}\tab \fs16\cf1\f0\charscalex100 {(a) The mode
l of the molecular envelope located}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f
0\charscalex100 {in the asymmetric unit of the}{ }\fs16\cf1\f0\charscalex100 {p3
}{ }\fs16\cf1\f0\charscalex100 {symmetry unit cell. The}\par\pard\li0\ri0\sl-200
\slmult0 \fs16\cf1\f0\charscalex100 {low-resolution shape and size of the envelo
pe is matched}\par\pard\li0\ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex100 {with
the form and size of the asymmetric unit. The}\par\pard\li0\ri0\sl-253\slmult0 \
fs16\cf1\f0\charscalex100 {parameters}{ }\fs16\cf1\f0\charscalex100 {R}\fs16\cf1
\f0\charscalex100 {,}{ }\fs16\cf1\f0\charscalex100 {a}\fs16\cf1\f0\charscalex100
{, and}{ }\fs16\cf1\f0\charscalex100 {b}{ }\fs16\cf1\f0\charscalex100 {define t
he origin and orientation of}\par\pard\li0\ri0\sl-186\slmult0 \fs16\cf1\f0\chars
calex100 {the molecule in the unit cell. (b,c) The normalized MFF}\par\pard\li0\
ri0\sl-226\slmult0 \fs16\cf1\f0\charscalex100 {calculated for 7-A}\fs16\cf1\f0\c

harscalex100 {\uc1\u730X }\fs16\cf1\f0\charscalex100 {resolution using discrete}


{\super \fs10\up3\cf1\f0\charscalex100 {33}}\fs16\cf1\f0\charscalex100 {and}\par
\pard\li0\ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex100 {continuous representati
on of bR, respectively. (d) The}\par\pard\li0\ri0\sl-226\slmult0 \fs16\cf1\f0\ch
arscalex100 {variation of the MFF of bR along the [1,0] direction}\par\pard\li0\
ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex100 {calculated for different resoluti
ons: 7 A}\fs16\cf1\f0\charscalex100 {\uc1\u730X }\fs16\cf1\f0\charscalex100 {(do
tted curve) and}\par\pard\li0\ri0\sl-226\slmult0 \fs16\cf1\f0\charscalex100 {5 A
}\fs16\cf1\f0\charscalex100 {\uc1\u730X }\fs16\cf1\f0\charscalex100 {(dashed cur
ve). The solid curve represents the case of}\par\pard\li0\ri0\sl-213\slmult0 \fs
16\cf1\f0\charscalex100 {the continuous representation of bR. [Color figure can
be}\par\pard\li0\ri0\sl-226\slmult0 \fs16\cf1\f0\charscalex100 {viewed in the on
line issue, which is available at}\par\pard\li0\ri0\sl-213\slmult0 \fs16\cf1\f0\
charscalex100 {wileyonlinelibrary.com.]}\par\pard\li0\ri0\sl-213\slmult0 \fs18\c
f0\par\pard\li0\ri0\sl-373\slmult0 \fs18\cf1\f0\charscalex100 {molecular envelop
e to be a solid ellipsoid, covering}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f
0\charscalex100 {the maximum possible area of the asymmetric unit}\par\pard\li0\
ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {without intersecting its boundari
es; this last require-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex1
00 {ment is introduced to avoid overlapping of molecules}\par\pard\li0\ri0\sl-24
0\slmult0 \fs18\cf1\f0\charscalex100 {under the appropriate symmetry operations,
Figure}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {1(a). In su
ch a representation, the origin and the ori-}\par\pard\li0\ri0\sl-240\slmult0 \f
s18\cf1\f0\charscalex100 {entation of the low-resolution shape of the molecule}\
par\pard\li0\ri0\sl-280\slmult0 \*\tx3466\*\tx3733\*\tx4373\fs18\cf1\f0\charscal
ex100 {are characterized by three parameters,}\tab \fs18\cf1\f0\charscalex100 {R
}\fs18\cf1\f0\charscalex100 {,}\tab \fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f0\c
harscalex100 {, and}\tab \fs18\cf1\f0\charscalex100 {b}\par\pard\li0\ri0\sl-200\
slmult0 \fs18\cf1\f0\charscalex100 {The first two parameters define the origin o
f the}\par\pard\li0\ri0\sl-280\slmult0 \*\tx2986\*\tx3213\*\tx3440\*\tx3720\*\tx
4333\fs18\cf1\f0\charscalex100 {molecule in the asymmetric unit,}\tab \fs18\cf1\
f0\charscalex100 {R}\tab \fs18\cf1\f0\charscalex100 {}\tab \fs18\cf1\f0\charscale
x100 {(}\fs18\cf1\f0\charscalex100 {R}\tab \fs18\cf1\f0\charscalex100 {cos(}\fs1
8\cf1\f0\charscalex100 {a}\fs18\cf1\f0\charscalex100 {),}\tab \fs18\cf1\f0\chars
calex100 {R}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {sin(}\f
s18\cf1\f0\charscalex100 {a}\fs18\cf1\f0\charscalex100 {)), and}{ }\fs18\cf1\f0\
charscalex100 {b}{ }\fs18\cf1\f0\charscalex100 {defines the orientation of the m
olecule.}\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf1\f0\charscalex100 {The 7-A}\f
s18\cf1\f0\charscalex100 {\uc1\u730X }\fs18\cf1\f0\charscalex100 {resolution map
s of the MFF calculated using}\par\pard\li1840\ri0\sl-213\slmult0 \fs18\cf0\par\
pard\li1840\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1840\ri0\sl-280\slmult0 \*\t
x4253\fs10\cf1\f0\charscalex100 {PROTEIN SCIENCE VOL 20:457\uc1\u8212X464}\tab \
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d\li5786\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li5786\ri0\sl-200\slmult0 \*\tx70
26\fs18\cf1\f0\charscalex100 {to the length,}\tab \fs18\cf1\f0\charscalex100 {D}
\fs18\cf1\f0\charscalex100 {, of the central chord of the molecu-}\par\pard\li57
86\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {lar envelope, which is paralle
l to the (}\fs16\cf1\f0\charscalex100 {h}\fs16\cf1\f0\charscalex100 {,}\fs16\cf1
\f0\charscalex100 {k}\fs16\cf1\f0\charscalex100 {) plane.}\par\pard\li5786\ri0\s
l-253\slmult0 \*\tx8720\*\tx9173\*\tx9880\fs16\cf1\f0\charscalex100 {The intensi
ty ratio of two peaks,}\tab \fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscale

x100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charscalex100 {1}}\fs16


\cf1\f0\charscalex100 {,}\tab \fs16\cf1\f0\charscalex100 {k}{\sub \fs10\cf1\f0\c
harscalex100 {1}}\fs16\cf1\f0\charscalex100 {) and}\tab \fs16\cf1\f0\charscalex1
00 {I}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf
1\f0\charscalex100 {2}}\fs16\cf1\f0\charscalex100 {,}\par\pard\li5786\ri0\sl-240
\slmult0 \fs18\cf1\f0\charscalex100 {k}{\sub \fs12\cf1\f0\charscalex100 {2}}\fs1
8\cf1\f0\charscalex100 {) can, therefore, be estimated as}\par\pard\sect\sectd\s
bknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols3\colno1\colw6626\col
sr-0\colno2\colw3213\colsr-0\colno3\colw1960\pard\li5786\ri0\sl-213\slmult0 \fs1
8\cf0\par\pard\li5786\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li5786\ri0\sl-213\sl
mult0 \fs18\cf0\par\pard\li5786\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li5786\ri0
\sl-213\slmult0 \fs18\cf1\f0\charscalex100 {where}\par\column\pard\li0\ri0\sl-21
3\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-306\slmult0 \fs18\cf1\f0\charscalex100 {
P}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {I h}{\sub \fs12\
cf1\f0\charscalex100 {1}}\fs18\cf1\f0\charscalex100 {;}{ }\fs18\cf1\f0\charscale
x100 {k}{\sub \fs12\cf1\f0\charscalex100 {1}}\fs18\cf1\f0\charscalex100 {}\fs18\c
f1\f0\charscalex100 {=}\fs18\cf1\f0\charscalex100 {I h}{\sub \fs12\cf1\f0\charsc
alex100 {2}}\fs18\cf1\f0\charscalex100 {;}{ }\fs18\cf1\f0\charscalex100 {k}{\sub
\fs12\cf1\f0\charscalex100 {2}}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charsc
alex100 {}\fs18\cf1\f0\charscalex100 {D}{\sub \fs12\cf1\f0\charscalex100 {1}}\fs1
8\cf1\f0\charscalex100 {=}\fs18\cf1\f0\charscalex100 {D}{\sub \fs12\cf1\f0\chars
calex100 {2}}\fs18\cf1\f0\charscalex100 {}\fs12\cf1\f0\charscalex100 {2}{\sub \fs
18\cf1\f0\charscalex100 {;}}\par\pard\li693\ri0\sl-333\slmult0 \fs18\cf1\f0\char
scalex100 {q}\fs12\cf1\f0\charscalex100 {\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc
1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259
X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u6
4259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc
1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259
X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u6
4259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc
1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X}\par\pard\li133\ri0\sl-240\slmult0
\*\tx866\*\tx1866\fs18\cf1\f0\charscalex100 {D}{\sub \fs12\cf1\f0\charscalex100
{1}}\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {2}\tab \fs18\cf1
\f0\charscalex100 {a}\fs12\cf1\f0\charscalex100 {2}{ }\fs18\cf1\f0\charscalex100
{cos}\fs12\cf1\f0\charscalex100 {2}\fs18\cf1\f0\charscalex100 { }\fs18\cf1\f0\cha
rscalex100 {c}\tab \fs18\cf1\f0\charscalex100 {b}\fs12\cf1\f0\charscalex100 {2}{
}\fs18\cf1\f0\charscalex100 {sin}{\super \fs12\up3\cf1\f0\charscalex100 {2}}\fs
18\cf1\f0\charscalex100 { }\fs18\cf1\f0\charscalex100 {c}{ }\fs18\cf1\f0\charscale
x100 {;}\par\column\pard\li200\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li200\ri0\s
l-280\slmult0 \fs18\cf1\f0\charscalex100 {(4)}\par\pard\li0\ri0\sl-213\slmult0 \
fs18\cf0\par\pard\li0\ri0\sl-373\slmult0 \fs18\cf1\f0\charscalex100 {D}{\sub \fs
12\cf1\f0\charscalex100 {2}}\fs18\cf1\f0\charscalex100 {}\par\pard\sect\sectd\sbk
none\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols2\colno1\colw5893\colsr
-0\colno2\colw5906\pard\li5786\ri0\sl-360\slmult0 \fs18\cf1\f0\charscalex100 {2}
\par\column\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {q}\fs12\cf1\
f0\charscalex100 {\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\u
c1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u6425
9X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u
64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\u
c1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u6425
9X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u
64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\u
c1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u6425
9X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u
64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X\uc1\u64259X}\par\pard\li173\ri0\sl-24
0\slmult0 \*\tx2693\*\tx2906\*\tx3333\*\tx3546\fs18\cf1\f0\charscalex100 {a}\fs1
2\cf1\f0\charscalex100 {2}{ }\fs18\cf1\f0\charscalex100 {cos}\fs12\cf1\f0\charsc
alex100 {2}\fs18\cf1\f0\charscalex100 {}\fs18\cf1\f0\charscalex100 {c}{ }\fs18\cf
1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {e}\fs18\cf1\f0\charscalex100
{ }{ }\fs18\cf1\f0\charscalex100 {b}\fs12\cf1\f0\charscalex100 {2}{ }\fs18\cf1\f0\

charscalex100 {sin}{\super \fs12\up3\cf1\f0\charscalex100 {2}}\fs18\cf1\f0\chars


calex100 {}\fs18\cf1\f0\charscalex100 {c}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\
cf1\f0\charscalex100 {e}\fs18\cf1\f0\charscalex100 {}\fs18\cf1\f0\charscalex100 {
,}\tab \fs18\cf1\f0\charscalex100 {a}\tab \fs18\cf1\f0\charscalex100 {and}\tab \
fs18\cf1\f0\charscalex100 {b}\tab \fs18\cf1\f0\charscalex100 {are main}\par\pard
\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols2\colno1
\colw5786\colsr-0\colno2\colw6013\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\p
ard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18
\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slm
ult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\
sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\l
i1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\
par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0
\fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-21
3\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013
\ri0\sl-226\slmult0 \*\tx1760\fs16\cf1\f0\charscalex100 {Figure 2.}\tab \fs16\cf
1\f0\charscalex100 {(a) The geometrical interpretation of Eq. (4). (b)}\par\pard
\li1013\ri0\sl-253\slmult0 \fs16\cf1\f0\charscalex100 {The variation of the}{ }\
fs16\cf1\f0\charscalex100 {c}{ }\fs16\cf1\f0\charscalex100 {parameter as a funct
ion of the intensity}\par\pard\li1013\ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex
100 {ratio of two diffraction peaks,}{ }\fs16\cf1\f0\charscalex100 {P}\fs16\cf1\
f0\charscalex100 {, calculated for three selected}\par\pard\li1013\ri0\sl-200\sl
mult0 \fs16\cf1\f0\charscalex100 {orientations of the molecular envelope: (black
asterisks) as}\par\pard\li1013\ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex100 {s
hown in Figure 2(a), (blue asterisks) rotated by 120}{ }\fs16\cf1\f0\charscalex1
00 {, and}\par\pard\li1013\ri0\sl-226\slmult0 \*\tx2946\fs16\cf1\f0\charscalex10
0 {(red asterisks) rotated by}\tab \fs16\cf1\f0\charscalex100 {120}{ }\fs16\cf1\
f0\charscalex100 {. The vertical lines}\par\pard\li1013\ri0\sl-240\slmult0 \fs16
\cf1\f0\charscalex100 {correspond to the intensity ratios}{ }\fs16\cf1\f0\charsc
alex100 {P}{ }\fs16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0\charscalex100 {1/2 and
}{ }\fs16\cf1\f0\charscalex100 {P}{ }\fs16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0
\charscalex100 {2/1. A, B,}\par\pard\li1013\ri0\sl-226\slmult0 \fs16\cf1\f0\char
scalex100 {C, and D indicate four possible solutions for}{ }\fs16\cf1\f0\charsca
lex100 {c}\fs16\cf1\f0\charscalex100 {,}\par\pard\li1013\ri0\sl-213\slmult0 \fs1
6\cf1\f0\charscalex100 {corresponding to}{ }\fs16\cf1\f0\charscalex100 {P}{ }\fs
16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0\charscalex100 {0.5. [Color figure can b
e viewed in the}\par\pard\li1013\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {
online issue, which is available at wileyonlinelibrary.com.]}\par\pard\li1013\ri
0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-280\slmult0 \fs18\cf1\f0\chars
calex100 {discrete}{\super \fs12\up3\cf1\f0\charscalex100 {33}}\fs18\cf1\f0\char
scalex100 {and continuous representations of the bR}\par\pard\li1013\ri0\sl-240\
slmult0 \fs16\cf1\f0\charscalex100 {molecule are shown in Figure 1(b,c), respect
ively.}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {Figure 1(
d) shows the variation of the MFF of the bR}\par\pard\li1013\ri0\sl-240\slmult0
\fs16\cf1\f0\charscalex100 {molecule}{\super \fs10\up3\cf1\f0\charscalex100 {33}
}\fs16\cf1\f0\charscalex100 {along the [1,0] crystallographic direction}\par\par
d\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {calculated for different
resolution limits. The solid}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\c
harscalex100 {curve represents the case of scattering from the mo-}\par\pard\li1
013\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {lecular envelope assuming a u
niform electron density}\par\pard\li1013\ri0\sl-240\slmult0 \fs16\cf1\f0\charsca
lex100 {distribution. It can be seen [Fig. 1(d)] that the MFF}\par\pard\li1013\r
i0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {calculated at different resolution
s are almost identi-}\par\pard\li1013\ri0\sl-240\slmult0 \*\tx3026\fs18\cf1\f0\c
harscalex100 {cal if the criterion SD}\tab \fs18\cf1\f0\charscalex100 {1 is sati
sfied.}\par\pard\li1373\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {For low-r
esolution structure analysis, therefore,}\par\pard\li1013\ri0\sl-240\slmult0 \fs
18\cf1\f0\charscalex100 {the orientation and origin of the molecule, rather}\par
\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {than the actual dist
ribution of electron density}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\ch

arscalex100 {inside the molecular envelope, is the dominant factor}\par\pard\li1


013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {affecting the amplitude of th
e structure factor. In}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscal
ex100 {this way, different positions and orientations of the}\par\pard\li1013\ri
0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {molecule give different ratios of i
ntensities for the}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex10
0 {pair of Bragg reflections.}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\
pard\li1013\ri0\sl-306\slmult0 \fs18\cf1\f0\charscalex100 {The uniqueness of the
solution}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {If we
consider the shape of the molecular envelope}\par\pard\li1013\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {to be an ellipsoid, then for the low-resolution lim
it}\par\pard\li1013\ri0\sl-280\slmult0 \*\tx4266\fs18\cf1\f0\charscalex100 {the
amplitude of the structure factor,}\tab \fs18\cf1\f0\charscalex100 {F}\fs18\cf1\
f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {S}\fs18\cf1\f0\charscalex100 {),
of a par-}\par\pard\li1013\ri0\sl-200\slmult0 \fs18\cf1\f0\charscalex100 {ticul
ar crystallographic plane (}\fs18\cf1\f0\charscalex100 {h}\fs18\cf1\f0\charscale
x100 {,}\fs18\cf1\f0\charscalex100 {k}\fs18\cf1\f0\charscalex100 {), is proporti
onal}\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\sl
mult0 \fs18\cf0\par\pard\li1013\ri0\sl-293\slmult0 \*\tx1506\fs18\cf1\f0\charsca
lex100 {460}\tab \fs12\cf1\f0\charscalex100 {PROTEINSCIENCE.ORG}\par\column\pard
\li0\ri0\sl-293\slmult0 \*\tx1906\*\tx2080\*\tx4360\fs18\cf1\f0\charscalex100 {a
xes of the ellipsoid,}\tab \fs18\cf1\f0\charscalex100 {c}\tab \fs18\cf1\f0\chars
calex100 {is the angle between the}\tab \fs18\cf1\f0\charscalex100 {a}\par\pard\
li0\ri0\sl-240\slmult0 \*\tx1160\*\tx2480\*\tx2653\fs18\cf1\f0\charscalex100 {an
d the (}\fs18\cf1\f0\charscalex100 {h}\fs12\cf1\f0\charscalex100 {1}\fs18\cf1\f0
\charscalex100 {,}\tab \fs18\cf1\f0\charscalex100 {k}\fs12\cf1\f0\charscalex100
{1}\fs18\cf1\f0\charscalex100 {) plane, and}\tab \fs18\cf1\f0\charscalex100 {e}\
tab \fs18\cf1\f0\charscalex100 {is the angle between}\par\pard\li0\ri0\sl-213\sl
mult0 \fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {h}{\sub \fs12\cf
1\f0\charscalex100 {1}}\fs18\cf1\f0\charscalex100 {,}{ }\fs18\cf1\f0\charscalex1
00 {k}{\sub \fs12\cf1\f0\charscalex100 {1}}\fs18\cf1\f0\charscalex100 {) and (}\
fs18\cf1\f0\charscalex100 {h}{\sub \fs12\cf1\f0\charscalex100 {2}}\fs18\cf1\f0\c
harscalex100 {,}{ }\fs18\cf1\f0\charscalex100 {k}{\sub \fs12\cf1\f0\charscalex10
0 {2}}\fs18\cf1\f0\charscalex100 {) planes, Figure 2(a). Then the ori-}\par\pard
\li0\ri0\sl-266\slmult0 \*\tx2266\fs18\cf1\f0\charscalex100 {entation of the env
elope,}\tab \fs18\cf1\f0\charscalex100 {c}\fs18\cf1\f0\charscalex100 {, can be c
alculated as a}\par\pard\li0\ri0\sl-240\slmult0 \*\tx3306\*\tx3546\*\tx4333\fs18
\cf1\f0\charscalex100 {function of the following parameters,}\tab \fs18\cf1\f0\c
harscalex100 {P}\fs18\cf1\f0\charscalex100 {,}\tab \fs18\cf1\f0\charscalex100 {b
}\fs18\cf1\f0\charscalex100 {/}\fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f0\charsc
alex100 {, and}\tab \fs18\cf1\f0\charscalex100 {e}\fs18\cf1\f0\charscalex100 {.}
\par\pard\li0\ri0\sl-240\slmult0 \*\tx2986\fs18\cf1\f0\charscalex100 {Figure 2(b
) shows the variation of}\tab \fs18\cf1\f0\charscalex100 {c}{ }\fs18\cf1\f0\char
scalex100 {as a function of}\par\pard\li0\ri0\sl-200\slmult0 \fs18\cf1\f0\charsc
alex100 {P}{ }\fs18\cf1\f0\charscalex100 {for three selected orientations of the
molecular en-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {velo
pe: (1) as shown in Figure 2(a), (2) rotated by}\par\pard\li0\ri0\sl-240\slmult0
\*\tx373\*\tx2253\*\tx2626\fs18\cf1\f0\charscalex100 {120}\tab \fs18\cf1\f0\cha
rscalex100 {, and (3) rotated by}\tab \fs18\cf1\f0\charscalex100 {120}\tab \fs18
\cf1\f0\charscalex100 {. The vertical lines in}\par\pard\li0\ri0\sl-253\slmult0
\*\tx3920\*\tx4320\fs18\cf1\f0\charscalex100 {Figure 2(b) correspond to the inte
nsity ratios}\tab \fs18\cf1\f0\charscalex100 {P}{ }\fs18\cf1\f0\charscalex100 {}\
tab \fs18\cf1\f0\charscalex100 {1/}\par\pard\li0\ri0\sl-240\slmult0 \*\tx573\*\t
x773\*\tx986\fs18\cf1\f0\charscalex100 {2 and}\tab \fs18\cf1\f0\charscalex100 {P
}\tab \fs18\cf1\f0\charscalex100 {}\tab \fs18\cf1\f0\charscalex100 {2/1. In this
example, we used the inten-}\par\pard\li0\ri0\sl-226\slmult0 \fs18\cf1\f0\charsc
alex100 {sity ratio,}{ }\fs18\cf1\f0\charscalex100 {P}\fs18\cf1\f0\charscalex100
{, of two diffraction peaks, (1,0) and (1,1).}\par\pard\li0\ri0\sl-240\slmult0
\fs18\cf1\f0\charscalex100 {As one can see, there are four possible solutions,}\
par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {marked A, B, C, and

D, corresponding to the}\par\pard\li0\ri0\sl-253\slmult0 \*\tx3253\*\tx3653\fs18


\cf1\f0\charscalex100 {selected values of the intensity ratio,}\tab \fs18\cf1\f0
\charscalex100 {P}{ }\fs18\cf1\f0\charscalex100 {}\tab \fs18\cf1\f0\charscalex100
{1/2. Pairs}\par\pard\li0\ri0\sl-226\slmult0 \fs18\cf1\f0\charscalex100 {of sol
utions (A&B) and (C&D) are separated by the}\par\pard\li0\ri0\sl-280\slmult0 \*\
tx546\*\tx853\*\tx1080\fs18\cf1\f0\charscalex100 {angle}\tab \fs18\cf1\f0\charsc
alex100 {Da}\tab \fs18\cf1\f0\charscalex100 {}\tab \fs18\cf1\f0\charscalex100 {60
}{ }\fs18\cf1\f0\charscalex100 {, and related to each other through}\par\pard\li
0\ri0\sl-200\slmult0 \fs18\cf1\f0\charscalex100 {the inversion operation. The ex
istence of (A and B)}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100
{or (C and D) pairs has a simple geometrical interpre-}\par\pard\li0\ri0\sl-253
\slmult0 \fs18\cf1\f0\charscalex100 {tation. Because of the symmetry of the enve
lope,}\par\pard\li0\ri0\sl-226\slmult0 \fs18\cf1\f0\charscalex100 {2mm}\fs18\cf1
\f0\charscalex100 {, two different orientations of the envelope give}\par\pard\l
i0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {exactly the same ratios of the
central chords and,}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100
{therefore, equal intensity ratios of the pair of reflec-}\par\pard\li0\ri0\sl240\slmult0 \fs18\cf1\f0\charscalex100 {tions. We may call these differently ori
ented enve-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {lopes t
he \uc1\u8216X\uc1\u8216Xcrystallographic twins.\uc1\u8217X\uc1\u8217X}\par\pard
\li373\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {We then calculated the var
iations of intensity}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100
{ratios of three pairs of reflections, (1,1) and (2,0), (2,2)}\par\pard\li0\ri0
\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {and (2,0), and (1,2) and (2,1) for t
he bR crystal, as func-}\par\pard\li0\ri0\sl-280\slmult0 \*\tx1173\fs18\cf1\f0\c
harscalex100 {tions of}{ }\fs18\cf1\f0\charscalex100 {Da}{ }\fs18\cf1\f0\charsca
lex100 {}\tab \fs18\cf1\f0\charscalex100 {a}\fs12\cf1\f0\charscalex100 {True}{\su
per \fs18\up0\cf1\f0\charscalex100 {}}\fs18\cf1\f0\charscalex100 {a}{ }\fs18\cf1\
f0\charscalex100 {and}{ }\fs18\cf1\f0\charscalex100 {Db}{ }\fs18\cf1\f0\charscal
ex100 {}{ }\fs18\cf1\f0\charscalex100 {b}\fs12\cf1\f0\charscalex100 {True}{\super
\fs18\up0\cf1\f0\charscalex100 {}}\fs18\cf1\f0\charscalex100 {b}\fs18\cf1\f0\cha
rscalex100 {, Figure 3.}\par\pard\li0\ri0\sl-240\slmult0 \*\tx1293\fs18\cf1\f0\c
harscalex100 {a}\fs12\cf1\f0\charscalex100 {True}{\super \fs18\cf1\f0\charscalex
100 {and}}\fs18\cf1\f0\charscalex100 {b}\fs12\cf1\f0\charscalex100 {True}\tab \f
s18\cf1\f0\charscalex100 {correspond to the exact location of bR}\par\pard\li0\r
i0\sl-240\slmult0 \*\tx3693\fs18\cf1\f0\charscalex100 {molecule in the asymmetri
c unit.}{\super \fs12\up3\cf1\f0\charscalex100 {33}}\fs18\cf1\f0\charscalex100 {
D}\fs18\cf1\f0\charscalex100 {P}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\c
harscalex100 {P}\tab \fs18\cf1\f0\charscalex100 {P}\fs12\cf1\f0\charscalex100 {T
rue}{ }\fs18\cf1\f0\charscalex100 {rep-}\par\pard\li0\ri0\sl-213\slmult0 \fs18\c
f1\f0\charscalex100 {resents the difference between the calculated values of}\pa
r\pard\li0\ri0\sl-226\slmult0 \fs18\cf1\f0\charscalex100 {intensity ratios and t
he exact values.}{\super \fs12\up3\cf1\f0\charscalex100 {33}}\fs18\cf1\f0\charsc
alex100 {For each pair}\par\pard\li0\ri0\sl-280\slmult0 \fs18\cf1\f0\charscalex1
00 {of reflections there are four possible values of}{ }\fs18\cf1\f0\charscalex1
00 {Da}{ }\fs18\cf1\f0\charscalex100 {when}\par\pard\li0\ri0\sl-240\slmult0 \*\t
x1760\fs18\cf1\f0\charscalex100 {D}\fs18\cf1\f0\charscalex100 {P}{ }\fs18\cf1\f0
\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {0. Only}{ }\fs18\cf1\f0\charscale
x100 {Da}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {0}\tab \f
s18\cf1\f0\charscalex100 {(the initial orientation),}{ }\fs18\cf1\f0\charscalex1
00 {Da}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {60}\par\par
d\li0\ri0\sl-240\slmult0 \*\tx3360\fs18\cf1\f0\charscalex100 {(the inversion ope
ration), and}{ }\fs18\cf1\f0\charscalex100 {Da}{ }\fs18\cf1\f0\charscalex100 {}{
}\fs18\cf1\f0\charscalex100 {120}\tab \fs18\cf1\f0\charscalex100 {(the threefold
}\par\pard\li0\ri0\sl-200\slmult0 \fs18\cf1\f0\charscalex100 {symmetry operation
) correspond to such an orientation}\par\pard\li0\ri0\sl-280\slmult0 \fs18\cf1\f
0\charscalex100 {of the molecular envelope when}{ }\fs18\cf1\f0\charscalex100 {D
}\fs18\cf1\f0\charscalex100 {P}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\ch
arscalex100 {0 for all pair of}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\cha
rscalex100 {reflections. Consequently,}{ }\fs18\cf1\f0\charscalex100 {D}\fs18\cf

1\f0\charscalex100 {P}{ }\fs18\cf1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex1


00 {0 for all pair of reflec-}\par\pard\li0\ri0\sl-240\slmult0 \*\tx1960\fs18\cf
1\f0\charscalex100 {tions only when}{ }\fs18\cf1\f0\charscalex100 {Db}{ }\fs18\c
f1\f0\charscalex100 {}{ }\fs18\cf1\f0\charscalex100 {0}\tab \fs18\cf1\f0\charscal
ex100 {.}\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmu
lt0 \fs18\cf0\par\pard\li0\ri0\sl-186\slmult0 \fs18\cf1\f0\charscalex100 {Decomp
osition of completely overlapping}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\
charscalex100 {reflections}\par\pard\li0\ri0\sl-280\slmult0 \*\tx3240\*\tx3733\*
\tx4000\fs18\cf1\f0\charscalex100 {A 2D molecular envelope function,}\tab \fs18\
cf1\f0\charscalex100 {E}\fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100
{R}\fs18\cf1\f0\charscalex100 {,}\tab \fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f
0\charscalex100 {,}\tab \fs18\cf1\f0\charscalex100 {b}\fs18\cf1\f0\charscalex100
{), is}\par\pard\li0\ri0\sl-200\slmult0 \fs18\cf1\f0\charscalex100 {defined to
describe the effective molecular bounda-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\
cf1\f0\charscalex100 {ries, Figure 1(a). We require that the total density}\par\
pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {inside the envelope is e
qual to the total number of}\par\pard\li533\ri0\sl-213\slmult0 \fs18\cf0\par\par
d\li533\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li533\ri0\sl-266\slmult0 \fs12\cf1
\f0\charscalex100 {A New Approach for Structure Analysis of 2D Membrane Proteins
}\par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\pa
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}}}}\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\margtsxn666\margbsxn173\par
d\li5786\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li5786\ri0\sl-200\slmult0 \*\tx83
86\*\tx8600\fs18\cf1\f0\charscalex100 {straints. If one can measure}\tab \fs18\c
f1\f0\charscalex100 {L}\tab \fs18\cf1\f0\charscalex100 {independent ratios,}\par
\pard\li5786\ri0\sl-280\slmult0 \*\tx8333\*\tx8480\*\tx8693\*\tx8933\*\tx9266\fs
16\cf1\f0\charscalex100 {which we label by the index}\tab \fs16\cf1\f0\charscale
x100 {j}\tab \fs16\cf1\f0\charscalex100 {}\tab \fs16\cf1\f0\charscalex100 {1,}\ta
b \fs16\cf1\f0\charscalex100 {. . .}\fs16\cf1\f0\charscalex100 {,}\tab \fs16\cf1
\f0\charscalex100 {L}\fs16\cf1\f0\charscalex100 {, then the}\par\pard\li5786\ri0
\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {mean discrepancy between the calcula
ted and the}\par\pard\li5786\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {meas
ured intensity ratios for these resolved reflec-}\par\pard\li5786\ri0\sl-240\slm
ult0 \fs18\cf1\f0\charscalex100 {tions can be calculated using}\par\pard\li9453\
ri0\sl-213\slmult0 \fs18\cf0\par\pard\li9453\ri0\sl-160\slmult0 \fs12\cf1\f0\cha
rscalex100 {2}\par\pard\li7586\ri0\sl-253\slmult0 \fs18\cf1\f0\charscalex100 {X}
{\super \fs12\up2\cf1\f0\charscalex100 {L}}\fs18\cf1\f0\charscalex100 {P}\fs12\c
f1\f0\charscalex100 {calc}{\sub \fs12\cf1\f0\charscalex100 {j}}\fs18\cf1\f0\char
scalex100 {}\fs18\cf1\f0\charscalex100 {R}\fs18\cf1\f0\charscalex100 {;}{ }\fs18\
cf1\f0\charscalex100 {a}\fs18\cf1\f0\charscalex100 {;}{ }\fs18\cf1\f0\charscalex
100 {b}\fs18\cf1\f0\charscalex100 { }{ }\fs18\cf1\f0\charscalex100 {P}{\sub \fs12
\cf1\f0\charscalex100 {j}}\par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\m
arglsxn666\margrsxn133\cols4\colno1\colw8600\colsr-0\colno2\colw933\colsr-0\coln
o3\colw506\colsr-0\colno4\colw1760\pard\li6440\ri0\sl-320\slmult0 \fs18\cf1\f0\c
harscalex100 {X}\fs18\cf1\f0\charscalex100 {}\fs18\cf1\f0\charscalex100 {R}\fs18\
cf1\f0\charscalex100 {;}{ }\fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f0\charscalex
100 {;}{ }\fs18\cf1\f0\charscalex100 {b}\fs18\cf1\f0\charscalex100 { }{\super \fs1
8\up4\cf1\f0\charscalex100 {1}}\par\pard\li7426\ri0\sl-226\slmult0 \fs18\cf1\f0\
charscalex100 {L}{\sub \fs12\cf1\f0\charscalex100 {j}}\fs12\cf1\f0\charscalex100
{}\fs12\cf1\f0\charscalex100 {1}\par\column{\trowd\trleft0\trkeep\trftsWidth1\tr
paddl0\trpaddr0\trpaddfl3\trpaddfr3 \clvertalt\clbrdrt\brdrnone\clbrdrl\brdrnone
\clbrdrb\brdrnone\clbrdrr\brdrnone\cltxlrtb\clNoWrap\clftsWidth3\clwWidth133\clp
adt0\clpadr0\clpadft3\clpadfr3\cellx9400 \pard\li0\ri0\sl-293\widctlpar\intbl \f
s18\cf1\f0\charscalex100 {r}\cell\clvertalt\clbrdrt\brdrnone\clbrdrl\brdrnone\cl
brdrb\brdrnone\clbrdrr\brdrnone\cltxlrtb\clNoWrap\clftsWidth3\clwWidth213\clpadt

0\clpadr0\clpadft3\clpadfr3\cellx9466 \pard\li0\ri0\sl-186\widctlpar\intbl \fs12


\cf1\f0\charscalex100 {2}\par\pard\li0\ri0\sl-120\widctlpar\intbl \fs12\cf1\f0\c
harscalex100 {j}\cell\row}\column\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\chars
calex100 {;}\par\column\pard\li0\ri0\sl-173\slmult0 \fs18\cf1\f0\charscalex100 {
(6)}\par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133
\cols2\colno1\colw5786\colsr-0\colno2\colw6013\pard\li1013\ri0\sl-213\slmult0 \f
s18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\
slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\r
i0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\par
d\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\c
f0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmul
t0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl
-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1
013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\pa
r\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \f
s18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\
slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\r
i0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\par
d\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\c
f0\par\pard\li1013\ri0\sl-346\slmult0 \*\tx1760\fs16\cf1\f0\charscalex100 {Figur
e 3.}\tab \fs16\cf1\f0\charscalex100 {Intensity ratios of three pairs of reflect
ions are}\par\pard\li1013\ri0\sl-266\slmult0 \*\tx4013\*\tx4386\fs16\cf1\f0\char
scalex100 {represented as functions of (a)}{ }\fs16\cf1\f0\charscalex100 {Da}{ }
\fs16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0\charscalex100 {a}\tab \fs16\cf1\f0\c
harscalex100 {a}\fs10\cf1\f0\charscalex100 {True}\tab \fs16\cf1\f0\charscalex100
{and (b)}{ }\fs16\cf1\f0\charscalex100 {Db}{ }\fs16\cf1\f0\charscalex100 {}\par\
pard\li1013\ri0\sl-213\slmult0 \*\tx1333\*\tx2533\*\tx2920\fs16\cf1\f0\charscale
x100 {b}\tab \fs16\cf1\f0\charscalex100 {b}\fs10\cf1\f0\charscalex100 {True}{\su
per \fs16\cf1\f0\charscalex100 {.}}\fs16\cf1\f0\charscalex100 {D}\fs16\cf1\f0\ch
arscalex100 {P}{ }\fs16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0\charscalex100 {P}\
tab \fs16\cf1\f0\charscalex100 {P}\fs10\cf1\f0\charscalex100 {True}\tab \fs16\cf
1\f0\charscalex100 {represents difference between}\par\pard\li1013\ri0\sl-186\sl
mult0 \fs16\cf1\f0\charscalex100 {the calculated values of intensity ratios and
the exact}\par\pard\li1013\ri0\sl-213\slmult0 \fs16\cf1\f0\charscalex100 {values
.}{\super \fs10\up2\cf1\f0\charscalex100 {33}}\fs16\cf1\f0\charscalex100 {[Color
figure can be viewed in the online issue,}\par\pard\li1013\ri0\sl-226\slmult0 \
fs16\cf1\f0\charscalex100 {which is available at wileyonlinelibrary.com.]}\par\p
ard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-213\slmult0 \fs18
\cf0\par\pard\li1013\ri0\sl-373\slmult0 \*\tx3253\*\tx4160\fs18\cf1\f0\charscale
x100 {electrons in bR, and that}\tab \fs18\cf1\f0\charscalex100 {E}\fs18\cf1\f0\
charscalex100 {(}\fs18\cf1\f0\charscalex100 {R}\fs18\cf1\f0\charscalex100 {,}{ }
\fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f0\charscalex100 {,}{ }\fs18\cf1\f0\char
scalex100 {b}\fs18\cf1\f0\charscalex100 {)}\tab \fs18\cf1\f0\charscalex100 {!}{
}\fs18\cf1\f0\charscalex100 {0 elsewhere.}\par\pard\li1013\ri0\sl-240\slmult0 \*
\tx2960\fs18\cf1\f0\charscalex100 {The structure factor,}\tab \fs18\cf1\f0\chars
calex100 {F}\fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {S}\fs18\cf
1\f0\charscalex100 {), for this approximation}\par\pard\li1013\ri0\sl-213\slmult
0 \fs18\cf1\f0\charscalex100 {can be rewritten as}\par\pard\li2546\ri0\sl-360\sl
mult0 \*\tx2813\*\tx3080\*\tx4266\fs18\cf1\f0\charscalex100 {n}\tab \fs18\cf1\f0
\charscalex100 {n}\tab \fs18\cf1\f0\charscalex100 {n}\tab \fs18\cf1\f0\charscale
x100 {o}\fs18\cf1\f0\charscalex100 {o}\fs18\cf1\f0\charscalex100 {o}\par\pard\li
1786\ri0\sl-293\slmult0 \*\tx2413\*\tx2666\*\tx2933\*\tx3200\*\tx3546\*\tx3746\*
\tx4613\*\tx5253\fs18\cf1\f0\charscalex100 {F}\fs18\cf1\f0\charscalex100 { }\fs18\
cf1\f0\charscalex100 {S}\tab \fs18\cf1\f0\charscalex100 {F}\fs18\cf1\f0\charscal
ex100 {^}\tab \fs18\cf1\f0\charscalex100 {U}\fs18\cf1\f0\charscalex100 {^}\tab \
fs18\cf1\f0\charscalex100 {G}\fs18\cf1\f0\charscalex100 {^}\tab \fs18\cf1\f0\cha
rscalex100 {T E R}\fs18\cf1\f0\charscalex100 {^}\tab \fs18\cf1\f0\charscalex100
{}\tab \fs18\cf1\f0\charscalex100 {;}{ }\fs18\cf1\f0\charscalex100 {a}\fs18\cf1\f
0\charscalex100 {;}{ }\fs18\cf1\f0\charscalex100 {b}\fs18\cf1\f0\charscalex100 {}
\fs18\cf1\f0\charscalex100 {g}\tab \fs18\cf1\f0\charscalex100 {;}\tab \fs18\cf1\

f0\charscalex100 {(5)}\par\pard\li1013\ri0\sl-426\slmult0 \*\tx1613\*\tx2026\fs1


6\cf1\f0\charscalex100 {where}\tab \fs16\cf1\f0\charscalex100 {F}\fs16\cf1\f0\ch
arscalex100 {^}\fs16\cf1\f0\charscalex100 {fg}\tab \fs16\cf1\f0\charscalex100 {d
enotes the Fourier transform operator,}\par\pard\li1013\ri0\sl-266\slmult0 \fs16
\cf1\f0\charscalex100 {U}\fs16\cf1\f0\charscalex100 {^}{ }\fs16\cf1\f0\charscale
x100 {fg}{ }\fs16\cf1\f0\charscalex100 {defines the translations of the unit cel
l across the}\par\pard\li1013\ri0\sl-266\slmult0 \fs18\cf1\f0\charscalex100 {cry
stallographic axis,}{ }\fs18\cf1\f0\charscalex100 {G}\fs18\cf1\f0\charscalex100
{^}{ }\fs18\cf1\f0\charscalex100 {fg}{ }\fs18\cf1\f0\charscalex100 {is the plane
group symme-}\par\column\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0
\sl-306\slmult0 \*\tx586\*\tx1200\fs18\cf1\f0\charscalex100 {where}\tab \fs18\cf
1\f0\charscalex100 {P}{\sub \fs12\cf1\f0\charscalex100 {j}}\fs18\cf1\f0\charscal
ex100 {and}\tab \fs18\cf1\f0\charscalex100 {P}\fs12\cf1\f0\charscalex100 {calc}{
\sub \fs12\cf1\f0\charscalex100 {j}}\fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\c
harscalex100 {R}\fs18\cf1\f0\charscalex100 {,}{ }\fs18\cf1\f0\charscalex100 {a}\
fs18\cf1\f0\charscalex100 {,}{ }\fs18\cf1\f0\charscalex100 {b}\fs18\cf1\f0\chars
calex100 {) are the}{ }\fs18\cf1\f0\charscalex100 {j}\fs18\cf1\f0\charscalex100
{th measured and}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {ca
lculated intensity ratios, respectively, and}{ }\fs16\cf1\f0\charscalex100 {r}{\
sub \fs10\cf1\f0\charscalex100 {j}}\fs16\cf1\f0\charscalex100 {is the}\par\pard\
li0\ri0\sl-213\slmult0 \fs18\cf1\f0\charscalex100 {standard deviation of the mea
surements,}{ }\fs18\cf1\f0\charscalex100 {P}{\sub \fs12\cf1\f0\charscalex100 {j}
}\fs18\cf1\f0\charscalex100 {.}\par\pard\li373\ri0\sl-226\slmult0 \fs16\cf1\f0\c
harscalex100 {The ability of the method to decompose com-}\par\pard\li0\ri0\sl-2
40\slmult0 \fs18\cf1\f0\charscalex100 {pletely overlapping reflections was first
tested with}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {the di
ffraction data simulated using the results pre-}\par\pard\li0\ri0\sl-240\slmult0
\fs16\cf1\f0\charscalex100 {sented in Ref. 33. The initial shape, the size, and
}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {the location of th
e molecular envelope were set as}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\c
harscalex100 {shown in Figure 1(a). The result of the analysis is}\par\pard\li0\
ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {shown in Tables I and II. The alg
orithm allows the}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {d
etermination of the values of intensity ratios of}\par\pard\li0\ri0\sl-240\slmul
t0 \fs16\cf1\f0\charscalex100 {completely overlapping reflections with a high le
vel}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {of accuracy. Fi
gure 4 shows the reconstructed and}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0
\charscalex100 {simulated MFF of bR. One can see that the magni-}\par\pard\li0\r
i0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {tudes and phases of calculated and
simulated MFF}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {are
in good agreement. It can also be seen from Ta-}\par\pard\li0\ri0\sl-240\slmult0
\fs16\cf1\f0\charscalex100 {ble II that the reconstructed and the simulated}\pa
r\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {phases of all reflecti
ons have identical signs and}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\chars
calex100 {comparable values.}\par\pard\li373\ri0\sl-240\slmult0 \fs16\cf1\f0\cha
rscalex100 {We then analyzed the powder diffraction pattern}\par\pard\li0\ri0\sl
-240\slmult0 \fs16\cf1\f0\charscalex100 {obtained from the PM crystal. The integ
rated inten-}\par\pard\li0\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {sities
of diffraction peaks up to a 4-A}\fs16\cf1\f0\charscalex100 {\uc1\u730X }\fs16\
cf1\f0\charscalex100 {resolution}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\c
harscalex100 {were estimated using the Le Bail analysis, shown in}\par\pard\li0\
ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {Figure 5(a) and Table III. The Le
Bail analysis was}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {
performed using RIETAN-2000.}{\super \fs12\up3\cf1\f0\charscalex100 {35}}\fs18\c
f1\f0\charscalex100 {The background of}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf
1\f0\charscalex100 {the powder diffraction pattern was represented by a}\par\par
d\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {composite background functi
on obtained with Pow-}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex10
0 {derX.}{\super \fs12\up3\cf1\f0\charscalex100 {36}}\fs18\cf1\f0\charscalex100
{The split pseudo-Voigt function of Toraya}{\super \fs12\up3\cf1\f0\charscalex10

0 {37}}\par\pard\li0\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {was used as


a profile function. The unit cell parame-}\par\pard\li0\ri0\sl-240\slmult0 \fs18
\cf1\f0\charscalex100 {ters of PM were determined using DICVOL.}{\super \fs12\up
3\cf1\f0\charscalex100 {38}}\fs18\cf1\f0\charscalex100 {The}\par\pard\li0\ri0\sl
-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-400\slmult0 \*\tx773\fs16\cf1\f0\char
scalex100 {Table I.}\tab \fs16\cf1\f0\charscalex100 {The Quality of the Decompos
ition of}\par\pard\li0\ri0\sl-173\slmult0 \fs16\cf1\f0\charscalex100 {Overlappin
g Reflections in Comparison with the}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\
f0\charscalex100 {Actual Data (Model)}{\super \fs10\up2\cf1\f0\charscalex100 {12
}}\par\pard\li280\ri0\sl-320\slmult0 \fs16\cf1\f0\charscalex100 {Intensity ratio
s of completely overlapping reflections}\par\pard\li2613\ri0\sl-320\slmult0 \fs1
6\cf1\f0\charscalex100 {P}{ }\fs16\cf1\f0\charscalex100 {}{ }\fs16\cf1\f0\charsca
lex100 {I}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs1
0\cf1\f0\charscalex100 {1}}\fs16\cf1\f0\charscalex100 {,}{ }\fs16\cf1\f0\charsca
lex100 {k}{\sub \fs10\cf1\f0\charscalex100 {1}}\fs16\cf1\f0\charscalex100 {)/}\f
s16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscale
x100 {h}{\sub \fs10\cf1\f0\charscalex100 {2}}\fs16\cf1\f0\charscalex100 {,}{ }\f
s16\cf1\f0\charscalex100 {k}{\sub \fs10\cf1\f0\charscalex100 {2}}\fs16\cf1\f0\ch
arscalex100 {)}\par\pard\li0\ri0\sl-280\slmult0 \*\tx1160\*\tx2320\*\tx4013\fs16
\cf1\f0\charscalex100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charsc
alex100 {1}}\fs16\cf1\f0\charscalex100 {,}{ }\fs16\cf1\f0\charscalex100 {k}{\sub
\fs10\cf1\f0\charscalex100 {1}}\fs16\cf1\f0\charscalex100 {)}\tab \fs16\cf1\f0\
charscalex100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charscalex100
{2}}\fs16\cf1\f0\charscalex100 {,}{ }\fs16\cf1\f0\charscalex100 {k}{\sub \fs10\c
f1\f0\charscalex100 {2}}\fs16\cf1\f0\charscalex100 {)}\tab \fs16\cf1\f0\charscal
ex100 {Reconstructed}\tab \fs14\cf1\f0\charscalex100 {Model}\par\pard\sect\sectd
\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols5\colno1\colw5786\c
olsr-0\colno2\colw1240\colsr-0\colno3\colw1453\colsr-0\colno4\colw1386\colsr-0\c
olno5\colw1933\pard\li1013\ri0\sl-266\slmult0 \*\tx2506\*\tx2906\fs18\cf1\f0\cha
rscalex100 {try operator, and}\tab \fs18\cf1\f0\charscalex100 {T}\fs18\cf1\f0\ch
arscalex100 {^}\fs18\cf1\f0\charscalex100 {fg}\tab \fs18\cf1\f0\charscalex100 {i
s the transformation operator,}\par\pard\li1013\ri0\sl-200\slmult0 \fs18\cf1\f0\
charscalex100 {which defines the orientation and origin of the mole-}\par\pard\l
i1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {cule in the asymmetric unit
. The intensity of the dif-}\par\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\cha
rscalex100 {fraction pattern can then be }\fs18\cf1\f0\charscalex100 {calculated
using Eq. (1).}\par\pard\li1373\ri0\sl-240\slmult0 \fs16\cf1\f0\charscalex100 {
Consider a pair of completely overlapping reflec-}\par\pard\li1013\ri0\sl-253\sl
mult0 \*\tx1546\*\tx2773\*\tx3226\*\tx4453\fs16\cf1\f0\charscalex100 {tions,}\ta
b \fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\chars
calex100 {h}{\sub \fs10\cf1\f0\charscalex100 {m}}\fs16\cf1\f0\charscalex100 {,}{
}\fs16\cf1\f0\charscalex100 {k}{\sub \fs10\cf1\f0\charscalex100 {m}}\fs16\cf1\f
0\charscalex100 {) and}\tab \fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscale
x100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charscalex100 {n}}\fs16
\cf1\f0\charscalex100 {,}\tab \fs16\cf1\f0\charscalex100 {k}{\sub \fs10\cf1\f0\c
harscalex100 {n}}\fs16\cf1\f0\charscalex100 {). The ratio}\tab \fs16\cf1\f0\char
scalex100 {P}{\sub \fs10\cf1\f0\charscalex100 {mn}}\fs16\cf1\f0\charscalex100 {}{
}\fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscalex100 {(}\fs16\cf1\f0\chars
calex100 {h}{\sub \fs10\cf1\f0\charscalex100 {m}}\fs16\cf1\f0\charscalex100 {,}\
par\pard\li1013\ri0\sl-240\slmult0 \*\tx1800\fs18\cf1\f0\charscalex100 {k}{\sub
\fs12\cf1\f0\charscalex100 {m}}\fs18\cf1\f0\charscalex100 {)/}\fs18\cf1\f0\chars
calex100 {I}\fs18\cf1\f0\charscalex100 {(}\fs18\cf1\f0\charscalex100 {h}{\sub \f
s12\cf1\f0\charscalex100 {n}}\fs18\cf1\f0\charscalex100 {,}\tab \fs18\cf1\f0\cha
rscalex100 {k}{\sub \fs12\cf1\f0\charscalex100 {n}}\fs18\cf1\f0\charscalex100 {)
can, therefore, be estimated by varying}\par\pard\li1013\ri0\sl-226\slmult0 \fs
16\cf1\f0\charscalex100 {the origin and the orientation of the approximated}\par
\pard\li1013\ri0\sl-240\slmult0 \fs18\cf1\f0\charscalex100 {molecular envelope a
nd using intensity ratios of}\par\pard\li1013\ri0\sl-253\slmult0 \*\tx2746\*\tx3
333\*\tx3800\*\tx4533\fs16\cf1\f0\charscalex100 {resolved reflections}\tab \fs16
\cf1\f0\charscalex100 {P}{\sub \fs10\cf1\f0\charscalex100 {pq}}\fs16\cf1\f0\char

scalex100 {}\tab \fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\charscalex100 {(}\fs1


6\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charscalex100 {p}}\fs16\cf1\f0\char
scalex100 {,}\tab \fs16\cf1\f0\charscalex100 {k}{\sub \fs10\cf1\f0\charscalex100
{p}}\fs16\cf1\f0\charscalex100 {)/}\fs16\cf1\f0\charscalex100 {I}\fs16\cf1\f0\c
harscalex100 {(}\fs16\cf1\f0\charscalex100 {h}{\sub \fs10\cf1\f0\charscalex100 {
q}}\fs16\cf1\f0\charscalex100 {,}\tab \fs16\cf1\f0\charscalex100 {k}{\sub \fs10\
cf1\f0\charscalex100 {q}}\fs16\cf1\f0\charscalex100 {) as con-}\par\column\pard\
li0\ri0\sl-253\slmult0 \fs16\cf1\f0\charscalex100 {(1, 2)}\par\pard\li0\ri0\sl-2
00\slmult0 \fs16\cf1\f0\charscalex100 {(1, 3)}\par\pard\li0\ri0\sl-200\slmult0 \
fs16\cf1\f0\charscalex100 {(2, 3)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\
charscalex100 {(1, 4)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex10
0 {(2, 4)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(1, 5)}\p
ar\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(3, 4)}\par\pard\li0\
ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(2, 5)}\par\pard\li0\ri0\sl-200\s
lmult0 \fs16\cf1\f0\charscalex100 {(1, 6)}\par\pard\li0\ri0\sl-200\slmult0 \fs16
\cf1\f0\charscalex100 {(2, 6)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\char
scalex100 {(1, 7)}\par\column\pard\li0\ri0\sl-253\slmult0 \fs16\cf1\f0\charscale
x100 {(2, 1)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(3, 1)
}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(3, 2)}\par\pard\l
i0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(4, 1)}\par\pard\li0\ri0\sl-20
0\slmult0 \fs16\cf1\f0\charscalex100 {(4, 2)}\par\pard\li0\ri0\sl-200\slmult0 \f
s16\cf1\f0\charscalex100 {(5, 1)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\c
harscalex100 {(4, 3)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100
{(5, 2)}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(6, 1)}\pa
r\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(6, 2)}\par\pard\li0\r
i0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {(7, 1)}\par\column\pard\li0\ri0\sl
-253\slmult0 \fs16\cf1\f0\charscalex100 {1.61}\par\pard\li0\ri0\sl-200\slmult0 \
fs16\cf1\f0\charscalex100 {0.73}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\ch
arscalex100 {0.60}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {1
.96}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {1.61}\par\pard\
li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {3.00}\par\pard\li0\ri0\sl-200
\slmult0 \fs16\cf1\f0\charscalex100 {0.16}\par\pard\li0\ri0\sl-200\slmult0 \fs16
\cf1\f0\charscalex100 {0.40}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charsc
alex100 {0.25}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {0.96}
\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {0.95}\par\column\pa
rd\li0\ri0\sl-253\slmult0 \fs16\cf1\f0\charscalex100 {2.11}\par\pard\li0\ri0\sl200\slmult0 \fs16\cf1\f0\charscalex100 {0.77}\par\pard\li0\ri0\sl-200\slmult0 \f
s16\cf1\f0\charscalex100 {0.47}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\cha
rscalex100 {1.95}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {1.
48}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {7.47}\par\pard\l
i0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {0.15}\par\pard\li0\ri0\sl-200\
slmult0 \fs16\cf1\f0\charscalex100 {0.98}\par\pard\li0\ri0\sl-200\slmult0 \fs16\
cf1\f0\charscalex100 {0.02}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charsca
lex100 {1.16}\par\pard\li0\ri0\sl-200\slmult0 \fs16\cf1\f0\charscalex100 {0.59}\
par\pard\sect\sectd\sbknone\pgwsxn12600\pghsxn16200\marglsxn666\margrsxn133\cols
2\colno1\colw7626\colsr-0\colno2\colw4173\pard\li1013\ri0\sl-213\slmult0 \fs18\c
f0\par\pard\li1013\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li1013\ri0\sl-253\slmul
t0 \fs12\cf1\f0\charscalex100 {Dilanian et al.}\par\column\pard\li0\ri0\sl-213\s
lmult0 \fs18\cf0\par\pard\li0\ri0\sl-213\slmult0 \fs18\cf0\par\pard\li0\ri0\sl-2
80\slmult0 \*\tx2413\fs12\cf1\f0\charscalex100 {PROTEIN SCIENCE VOL 20:457\uc1\u
8212X464}\tab \fs18\cf1\f0\charscalex100 {461}\par}

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