Structure of Bacterial Outer Membrane Proteins

Bhaskar Ganguly Ph.D. Scholar Animal Biotechnology Laboratory College of Veterinary & Animal Sciences G.B.P.U.A. & T., Pantnagar INDIA - 263145

INTRODUCTION
20 % of all known proteins are found in membranes
Consist of transmembrane - helices; characterized by 20 - 30 non-polar a.a. with a predominance of aliphatic side chains at the center and aromatic residues at both the ends

Introduction

Outer membranes of Gram Negative Bacteria lack helical proteins.

Instead, - barrel proteins are present in outer membranes; characterized by an alternating sequence of polar & non-polar residues.
Contain all-next-neighbor antiparallel - sheets. However, - barrel proteins are absent from the outer membranes of mitochondrion and chloroplasts.

Polar residues Non-polar residues

STRUCTURE OF OMPs
Determined from X-ray crystallographic studies Requires 3-D crystals of purified proteins Difficulties: Limitations on the amount of recombinant OMPs that can be incorporated within a fixed membrane volume Altered membrane properties may be deleterious to the host Solution: Cytosolic expression of the OMP and reconstitution into miscelles

STRUCTURAL FEATURES OF OMPs

n {no. of strands} S {Shear number} R {radius} {tilt angle}

R = [(Sa)2 + (nb)2]0.5 / 2 tan = Sa / nb R = nb / 2 cos

a = 3.3 ; b = 4.4

n = 16

S = 14
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14.

2R

Structural features of OMPs

n S n+4 n is always even (n = 8 barrels are common) S is generally positive and always even - strands are anti-parallel and connected to next neighbor - barrel surface bears aliphatic side chains; rims of the barrel are lined by aromatic side chains

FUNCTIONS OF OMPs
OmpX: Stress modulator; binds foreign proteins OmpT: protease; role in pathogenicity OmpLA: phospholipase; forms channels in outer membrane for secretion of colicins and virulence factors Porins: e.g. OmpF, PhoE, OmpC, etc. {trimers consisting of three parallel - barrels}
OmpF osmoporin, low salt concentration PhoE phosphate uptake OmpC osmoporin, high salt concentration

Functions of OMPs
Maltoporin: Glucose oligomers Sucrose porin: Sucrose transport FhuA, FepA: iron transporters; interact with TonB of inner membrane that draws energy from cytosolic ATP

PORINS
Passive diffusion channels Large pores are guarded by oppositely charged residues at opposite sides
-

+
+

+ +

Disallows large non-polar molecules (viz. antibiotics) but allow polar molecules like sugars 3 classes: n = 16, n = 18, n = 22

PORINS
n = 16, S = 20, nearly circular cross-section, commonly associate to form trimers, pore size 10 , allow molecules of about 600 Da
Hydrophobic interface behaves like a cytoplasmic - Barrel

n = 18, trimeric, smaller cross-section than n = 18 porins, high selectivity, pore size 6 n = 22, iron transporters, monomeric, very large crosssection, filled with N-terminal 150 residue domain

STABILITY STUDIES
- Barrel is a robust and stable structure Large external loops present in - Barrels of OmpA were replaced by short-cuts The deletion mutants were functionally inactive; lacked biological functions in Fconjugation and as a receptor for Bacteriophages However, the trans-membrane - Barrel structure was retained

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