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. 1981 Apr;78(4):2302–2306. doi: 10.1073/pnas.78.4.2302

Matrix protein in planar membranes: clusters of channels in a native environment and their functional reassembly.

H Schindler, J P Rosenbusch
PMCID: PMC319333  PMID: 6264473

Abstract

Planar bilayers formed from Escherichia coli outer membrane vesicles exhibit conductance properties similar to those previously observed in bilayers reconstituted from aggregates of matrix protein, the major outer membrane protein. Discrete conductance steps are observed, reflecting voltage-dependent transmembrane channels. These exist in clusters which are activated by voltage. After activation, channels close with increasing potentials and reopen reversibly at lower voltage. Depending on the sign of the potential, two distinct closed states of the pores are observed. Cooperative interactions, hysteresis effects, relaxation times, and values of channel conductance depend on cluster size. These properties provide the reference data for the reconstitution of membrane function from individual components. Planar bilayers were formed from vesicles containing either solubilized matrix protein in a homogeneous trimeric state or bacterial glycolipid (lipopolysaccharide), or both. Activation of channel conductance required the presence of glycolipid and the formation of channel clusters, leading to conductance properties of the channels closely resembling those observed in native outer membranes. At very low concentrations of trimers, irreversible association to clusters by lateral diffusion was observed. Nearly quantitative recoveries of channels allowed the assignment of three pores per trimer.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bavoil P., Nikaido H., von Meyenburg K. Pleiotropic transport mutants of Escherichia coli lack porin, a major outer membrane protein. Mol Gen Genet. 1977 Dec 14;158(1):23–33. doi: 10.1007/BF00455116. [DOI] [PubMed] [Google Scholar]
  2. Benz R., Janko K., Boos W., Läuger P. Formation of large, ion-permeable membrane channels by the matrix protein (porin) of Escherichia coli. Biochim Biophys Acta. 1978 Aug 17;511(3):305–319. doi: 10.1016/0005-2736(78)90269-9. [DOI] [PubMed] [Google Scholar]
  3. Cherry R. J. Rotational and lateral diffusion of membrane proteins. Biochim Biophys Acta. 1979 Dec 20;559(4):289–327. doi: 10.1016/0304-4157(79)90009-1. [DOI] [PubMed] [Google Scholar]
  4. Ehrenstein G., Lecar H. Electrically gated ionic channels in lipid bilayers. Q Rev Biophys. 1977 Feb;10(1):1–34. doi: 10.1017/s0033583500000123. [DOI] [PubMed] [Google Scholar]
  5. Fried V. A., Rothfield L. I. Interactions between lipopolysaccharide and phosphatidylethanolamine in molecular monolayers. Biochim Biophys Acta. 1978 Dec 4;514(1):69–82. doi: 10.1016/0005-2736(78)90077-9. [DOI] [PubMed] [Google Scholar]
  6. Garavito R. M., Rosenbusch J. P. Three-dimensional crystals of an integral membrane protein: an initial x-ray analysis. J Cell Biol. 1980 Jul;86(1):327–329. doi: 10.1083/jcb.86.1.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Montal M., Mueller P. Formation of bimolecular membranes from lipid monolayers and a study of their electrical properties. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3561–3566. doi: 10.1073/pnas.69.12.3561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nakae T., Ishii J., Tokunaga M. Subunit structure of functional porin oligomers that form permeability channels in the other membrane of Escherichia coli. J Biol Chem. 1979 Mar 10;254(5):1457–1461. [PubMed] [Google Scholar]
  9. Nakae T. Outer membrane of Salmonella typhimurium: reconstitution of sucrose-permeable membrane vesicles. Biochem Biophys Res Commun. 1975 Jun 16;64(4):1224–1230. doi: 10.1016/0006-291x(75)90823-2. [DOI] [PubMed] [Google Scholar]
  10. Nikaido H., Nakae T. The outer membrane of Gram-negative bacteria. Adv Microb Physiol. 1979;20:163–250. doi: 10.1016/s0065-2911(08)60208-8. [DOI] [PubMed] [Google Scholar]
  11. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  12. Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
  13. Schindler H. Exchange and interactions between lipid layers at the surface of a liposome solution. Biochim Biophys Acta. 1979 Aug 7;555(2):316–336. doi: 10.1016/0005-2736(79)90171-8. [DOI] [PubMed] [Google Scholar]
  14. Schindler H. Formation of planar bilayers from artificial or native membrane vesicles. FEBS Lett. 1980 Dec 15;122(1):77–79. doi: 10.1016/0014-5793(80)80405-4. [DOI] [PubMed] [Google Scholar]
  15. Schindler H., Quast U. Functional acetylcholine receptor from Torpedo marmorata in planar membranes. Proc Natl Acad Sci U S A. 1980 May;77(5):3052–3056. doi: 10.1073/pnas.77.5.3052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schindler H., Rosenbusch J. P. Matrix protein from Escherichia coli outer membranes forms voltage-controlled channels in lipid bilayers. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3751–3755. doi: 10.1073/pnas.75.8.3751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schindler M., Osborn M. J., Koppel D. E. Lateral mobility in reconstituted membranes--comparisons with diffusion in polymers. Nature. 1980 Jan 24;283(5745):346–350. doi: 10.1038/283346a0. [DOI] [PubMed] [Google Scholar]
  18. Stankowski S., Gruenewald B. Evaluation of cooperativity for phase transitions in two- and three-dimensional systems. Biophys Chem. 1980 Oct;12(2):167–176. doi: 10.1016/0301-4622(80)80049-4. [DOI] [PubMed] [Google Scholar]
  19. Steven A. C., Heggeler B., Müller R., Kistler J., Rosenbusch J. P. Ultrastructure of a periodic protein layer in the outer membrane of Escherichia coli. J Cell Biol. 1977 Feb;72(2):292–301. doi: 10.1083/jcb.72.2.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ueki T., Mitsui T., Nikaido H. X-ray diffraction studies of outer membranes of Salmonella typhimurium. J Biochem. 1979 Jan;85(1):173–182. doi: 10.1093/oxfordjournals.jbchem.a132307. [DOI] [PubMed] [Google Scholar]
  21. Yamada H., Mizushima S. Interaction between major outer membrane protein (O-8) and lipopolysaccharide in Escherichia coli K12. Eur J Biochem. 1980 Jan;103(1):209–218. doi: 10.1111/j.1432-1033.1980.tb04305.x. [DOI] [PubMed] [Google Scholar]

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