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Escherichia Coli and Other Gram-Negative Bacteria Biochimica et Biophysica A cta, 737 (1983) 51 - 115 51 Elsevier Biomedical Press BBA 85241 MOLECULAR ARCHITECTURE AND FUNCTIONING OF THE OUTER MEMBRANE OF ESCHERICHIA COLI AND OTHER GRAM-NEGATIVE BACTERIA BEN LUGTENBERG a,, and LOEK VAN ALPHEN h " Department of Molecular Cell Biology' and Institute for Molecular Biology', State University, Transitorium 3, Padualaan 8, 3584 CH Utrecht and h Laboratorium voor de Gezondheidsleer, University of Amsterdam, Mauritskade 57, 1092 AD Amsterdam (The Netherlands) (Received July 26th, 1982) Contents Introduction ............................................................................. 52 A. Scope of this review ...................................................................... 52 B. Ecological considerations relevant to structure and functioning of the outer membrane of Enterobacteriaceae ........ 53 C. General description of the cell envelope of Gram-negative bacteria ..................................... 53 II. Methods for the isolation of outer membranes ...................................................... 58 A. E. coli and S. typhimurium ................................................................. 58 1. Isolation of peptidoglycan-less outer membranes after spheroplast formation ............................ 58 2. Isolation of outer membrane-peptidoglycan complexes ........................................... 58 3. Differential membrane solubilization using detergents ............................................ 59 4. Membrane separation based on charge differences of vesicles ....................................... 59 B. Other organisms ........................................................................ 59 II1. Individual constituents of the outer membrane ..................................................... 59 A. Composition of the outer membrane .......................................................... 59 B. Phospholipid .......................................................................... 60 C. Lipopolysaccharide ...................................................................... 61 1. Introduction ......................................................................... 61 2, Methods of isolation and purification ....................................................... 61 3. Chemical structure of lipopolysaccharides .................................................... 62 4. Effects of polymixin, EDTA and divalent cations ............................................... 64 D. Enterobacterial common antigen (ECA) ........................................................ 64 E. Proteins .............................................................................. 64 1. Introductory remarks ................................................................... 64 2. Enzymes in the outer membrane ........................................................... 67 3. Lipoproteins ......................................................................... 68 4. OmpA protein ....................................................................... 70 5. The family of peptidoglycan-associated general diffusion pore proteins ................................ 71 a. Introduction ....................................................................... 71 b. Function of general pore proteins ........................................................ 72 c. Purification and properties of general diffusion pore proteins ..................................... 75 d. Characteristics of individual general diffusion pore proteins ...................................... 76 * To whom correspondence should be addressed. heptose; KDO, 2-keto-3-deoxy-octulosonic acid; LPS, lipopoly- Abbreviations: Abe, abequose; ECA, Enterobacterial Common saccharide; NMR, nuclear magnetic resonance; PAL, pepti- Antigen; ESR, electron spin resonance; GIcN, glucosamine; doglycan-associated lipoprotein; Rha, rhamnose; SDS, sodium GIcNAc, N-acetyl-D-glucosamine; Hep. t-glycero-D-manno dodecyl sulphate. 0304-4157/83/0000-0000/$03.00 © 1983 Elsevier Science Publishers 52 i. OmpC protein and OmpF protein ..................................................... 76 ii. PhoE protein .................................................................... 76 iii. Salmonella pore proteins ............................................................ 77 iv. Other general diffusion pore proteins ................................................... 77 6, Characteristics of E. coil pore proteins not antigenically related to the family of peptidoglycan-associated general diffusion pore proteins .................................................................. 78 a. Bacteriophage T6 receptor protein ....................................................... 78 b. Bacteriophage lambda receptor protein .................................................... 78 c. Outer membrane protein involved in the uptake of vitamin B12 ................................... 80 d. Outer membrane proteins involved in the uptake of ferric ions .................................... 80 Characteristics of E. coil outer membrane proteins without identified function ........................... 80 a. Protein a ......................................................................... 80 b. Protein II1 ........................................................................ 81 c. LPS binding protein ................................................................. 81 d. Outer membrane proteins induced by sulphate limitation ....................................... 81 e. Phage- and plasmid-coded outer membrane proteins ........................................... 82 IV. Molecular organization of the outer membrane ..................................................... 82 A. Introduction ........................................................................... 82 B. Methods used for studying the localization of individual outer membrane constituents and their interactions ........ 84 1. Localization at the cell surface ............................................................ 84 2. Protein-protein and protein-peptidoglycan nearest neighbour associations .............................. 85 3 Interactions between individual proteins and LPS ............................................... 86 4. The lipid matrix and interactions of proteins with lipids .......................................... 86 C. Localization of LPS ...................................................................... 86 D. Localization of phospholipids ............................................................... 86 E. Localization of ECA ..................................................................... 87 F. Localization and topography of outer membrane proteins ........................................... 87 1. Introduction ......................................................................... 87 2. The major lipoprotein .................................................................. 88 3. OmpA protein ....................................................................... 89 4. Peptidoglycan-associated pore proteins ...................................................... 90 5. Are matrix (pore) proteins associated with peptidoglycan in vivo? .................................... 91 G. The lipid matrix ........................................................................ 93 1. Is the outermembrane a lipid bi,layer? ....................................................... 93 2. Nature of OM particles and OM pits on the fracture faces of the outer membrane observed with freeze-fracture electron microscopy .................................................................... 95 3. Physical properties of LPS and phospholipids in the outer membrane ................................. 97 4. Interactions of proteins with the lipid matrix .................................................. 99 5. Distribution of outer membrane constituents over both monolayers .................................. 100 H. Molecular organization of the outer membrane of Enterobacteriaceae ................................... 101 1. Outer membrane of other Gram-negative bacteria ................................................. 103 V. Future prospects .......................................................................... 103 Acknowledgements ............................................................................ 104 References .................................................................................. 104 I. Introduction tive and Gram-negative bacteria differ fundamen- tally with respect to the composition of their cell IA. Scope of this review walls. One of the major differences is that Gram- negative cells contain an outer membrane, located The cytosol of a bacterial cell is surrounded by at the outside of a monolayer of peptidoglycan. a complex cell envelope which usually consists of a This outer membrane forms the physical and func- cytoplasmic membrane and a cell wall. Gram-posi- tional barrier between the inside of the cell and its 53 environment. After methods for the isolation of Donor cells of several bacterial species can outer membranes became available, its composi- transfer (part of) their genetic information to tion, structure, function and biogenesis have been acceptor cells, the latter ones usually being related studied extensively in the last decade, especially in to the donor. Conjugative transfer, especially of the enteric bacteria Escherichia coli and Salmonella plasmid DNA, usually is the means by which typhimurium. Recently, a monograph [1] as well as genetic information coding for production
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