TRANSITION STATES-ANALOG

INTRODUCTION TO TRANSITION STATE

All chemical transformations transition state

In enzymatic study transition state is

central point to understanding catalysis

For catalysis to be efficient, the enzyme must

bind the transition-state more tightly than the substra

Transition states have short lifetimes

(near 10-13 detik) cannot be captured
or directly observed
INTRODUCTION TO TRANSITION STATE
Some approaches to understanding Enzyme-transit
state complementary
Structure activity correlations
Transition state analogs
Catalytic antibodies
Site directed mutagenasis
 TRANSITION STATE ANALOG

Stable molecule designed to resemble transition state

Should bind to an enzyme more tightly than does the

substrate in the ES complex 102-106 times more

Transition state analogs as very good inhibitors

design new drug

Limitation can not perfectly mimic transition state

Experimentally approaches to enzyme
transition state analog design

TS structure prediction
(KIE & computational approaches)

TS analogues design

Confirming TS
(Crystalography and Spectroscopy)

V.L. Schramm / Archives of Biochemistry and Biophysics 433 (2005)

 Proline racemase

Best inhibitors are the planar transition state analogs 160 x stronger

The tetrahedral substrate analog is a poor inhibitor (binds very weakly).

Lysozyme

Initial rates of lysozyme-calalyzed lysis of M.

lysodeikticus in the presence of inhibit,ors. A
mixture of TACA and TACL (X), TACA (tetra-N
acetylchitotetraose) (), GlcNAc(0).
 Purine nucleoside phosphorylase
(PNP) (dGuo (inosine) + PO4 guanine (hypoxanthine) + 2-deoxy-a-D-
ribose 1-PO4)

Immucilin H Transition state analog

Crystal structures of bovine purine nucleoside phosphorylase (PNP) with
substrate analogues (A), transition state analogue (B) and products (C)
bound at the catalytic sites. Conversion from substrate to transition
state analogue forms new hydrogen bonds and closer atomic contacts
(red in B) and one relaxed distance that contributes to the
nucleophilicity of phosphate (blue in B). In the product complex,
multiple atomic distances relax to form weaker contacts (blue in C)
 Purine nucleoside phosphorylase

the electrostatic potential similarity between the

transition state and the transition state inhibitor is
greater than between the substrate and the
transition state
 AMP nucleosidase

AMP + H2O adenine + ribose 5-phosphate

formycin A 50-phosphate binds >103 times

tighter to AMP nucleosidases than does
substrate
 AMP nucleosidase

The molecular electrostatic potential at the van der

Waals surface revealed similar electronic
structures of inhibitor and transition state because
of the common protonation at N7, which is not
present in the substrate
 HIV protease inhibitor

HIV protease inhibitor a molecule that mimics

the functionality and shape of the gaG protein
cleavage region O R
H
N Peptidebond
N
H
R O

R
O- O
H
N Transitionstateduring
N peptidebondhydrolysis
H
R O

OH R
H H
N N Transitionstateanalog:
hydroxyethylamine
R O
 Thank you

ENJOY
BIOCHEMISTRY