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. 1980 Jul;143(1):349–354. doi: 10.1128/jb.143.1.349-354.1980

Isolation and partial characterization of the major outer membrane protein of Chromatium vinosum.

B C Lane, R E Hurlbert
PMCID: PMC294244  PMID: 7400096

Abstract

The 42,000 major outer membrane protein of Chromatium vinosum was purified by a combination on ion-exchange chromatography, gel filtration, and isoelectric focusing. Upon isoelectric focusing, the final material produced four major hands. Three of the four bands were isolated and analyzed for similarity or differences. Protease peptide maps and cyanogen bromide maps of the three isoelectric species were identical. When the isolated isoelectric species were refocused, each produced multiple isoelectric species, suggesting that the procedure used was generating the multiple charged species. Protease treatment of the isolated outer membrane produced a 31,000 fragment from the 42,000 protein. This fragment was isolated by preparative sodium sulfate-polyacrylamide gel electrophoresis. Although the amino acid compositions of the 42,000 protein and its 31,000 trypsin fragment were different, their polarity index was the same (45%). The amino-terminal sequences of the 42,000 protein and 31,000 trypsin fragment were identical, and it concluded that the amino-terminal was buried in the membrane.

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Selected References

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

  1. Capaldi R. A., Vanderkooi G. The low polarity of many membrane proteins. Proc Natl Acad Sci U S A. 1972 Apr;69(4):930–932. doi: 10.1073/pnas.69.4.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  3. DiRienzo J. M., Nakamura K., Inouye M. The outer membrane proteins of Gram-negative bacteria: biosynthesis, assembly, and functions. Annu Rev Biochem. 1978;47:481–532. doi: 10.1146/annurev.bi.47.070178.002405. [DOI] [PubMed] [Google Scholar]
  4. Garten W., Henning U. Cell envelope and shape of Escherichia coli K12. Isolation and preliminary characterization of the major ghost-membrane proteins. Eur J Biochem. 1974 Sep 1;47(2):343–352. doi: 10.1111/j.1432-1033.1974.tb03699.x. [DOI] [PubMed] [Google Scholar]
  5. Garten W., Hindennach I., Henning U. The major proteins of the Escherichia coli outer cell envelope membrane. Characterization of proteins II* and III, comparison of all proteins. Eur J Biochem. 1975 Nov 1;59(1):215–221. doi: 10.1111/j.1432-1033.1975.tb02444.x. [DOI] [PubMed] [Google Scholar]
  6. HURLBERT R. E., LASCELLES J. RIBULOSE DIPHOSPHATE CARBOXYLASE IN THIORHODACEAE. J Gen Microbiol. 1963 Dec;33:445–458. doi: 10.1099/00221287-33-3-445. [DOI] [PubMed] [Google Scholar]
  7. Hasin M., Rottem S., Razin S. The outer membrane of Proteus mirabilis. I. Isolation and characterization of the outer and cytoplasmic membrane fractions. Biochim Biophys Acta. 1975 Feb 14;375(3):381–394. doi: 10.1016/0005-2736(75)90354-5. [DOI] [PubMed] [Google Scholar]
  8. Henning U., Sonntag I., Hindennach I. Mutants (ompA) affecting a major outer membrane protein of Escherichia coli K12. Eur J Biochem. 1978 Dec;92(2):491–498. doi: 10.1111/j.1432-1033.1978.tb12771.x. [DOI] [PubMed] [Google Scholar]
  9. Hui K. M., Hurlbert R. E. Modifiable chromatophore proteins in photosynthetic bacteria. J Bacteriol. 1979 Apr;138(1):207–217. doi: 10.1128/jb.138.1.207-217.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hurlbert R. E., Hurlbert I. M. Biological and physicochemical properties of the lipopolysaccharide of Chromatium vinosum. Infect Immun. 1977 Jun;16(3):983–994. doi: 10.1128/iai.16.3.983-994.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hurlbert R. E. Isolation and characterization of Chromatium vinosum membranes. Arch Microbiol. 1974;101(2):169–186. doi: 10.1007/BF00455937. [DOI] [PubMed] [Google Scholar]
  12. Johnston K. H., Gotschlich E. C. Isolation and characterization of the outer membrane of Neisseria gonorrhoeae. J Bacteriol. 1974 Jul;119(1):250–257. doi: 10.1128/jb.119.1.250-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lane B. C., Hurlbert R. E. Characterization of the cell wall and cell wall proteins of Chromatium vinosum. J Bacteriol. 1980 Mar;141(3):1386–1398. doi: 10.1128/jb.141.3.1386-1398.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lee D. R., Schnaitman C. A., Pugsley A. P. Chemical heterogeneity of major outer membrane pore proteins of Escherichia coli. J Bacteriol. 1979 Jun;138(3):861–870. doi: 10.1128/jb.138.3.861-870.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mizuno T., Kageyama M. Isolation and characterization of major outer membrane proteins of Pseudomonas aeruginosa strain PAO with special reference to peptidoglycan-associated protein. J Biochem. 1979 Oct;86(4):979–989. doi: 10.1093/oxfordjournals.jbchem.a132630. [DOI] [PubMed] [Google Scholar]
  16. Nakae T. Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. J Biol Chem. 1976 Apr 10;251(7):2176–2178. [PubMed] [Google Scholar]
  17. Nixdorff K., Fitzer H., Gmeiner J., Martin H. H. Reconstitution of model membranes from phospholipid and outer membrane proteins of Proteus mirabilis. Role of proteins in the formation of hydrophilic pores and protection of membranes against detergents. Eur J Biochem. 1977 Nov 15;81(1):63–69. doi: 10.1111/j.1432-1033.1977.tb11927.x. [DOI] [PubMed] [Google Scholar]
  18. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  19. Schnaitman C. A. Outer membrane proteins of Escherichia coli. 3. Evidence that the major protein of Escherichia coli O111 outer membrane consists of four distinct polypeptide species. J Bacteriol. 1974 May;118(2):442–453. doi: 10.1128/jb.118.2.442-453.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schnaitman C. A. Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J Bacteriol. 1970 Nov;104(2):890–901. doi: 10.1128/jb.104.2.890-901.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schweizer M., Hindennach I., Garten W., Henning U. Major proteins of the Escherichia coli outer cell envelope membrane. Interaction of protein II with lipopolysaccharide. Eur J Biochem. 1978 Jan 2;82(1):211–217. doi: 10.1111/j.1432-1033.1978.tb12013.x. [DOI] [PubMed] [Google Scholar]
  22. Uemura J., Mizushima S. Isolation of outer membrane proteins of Escherchia coli and their characterization on polyacrylamide gel. Biochim Biophys Acta. 1975 Dec 1;413(2):163–176. doi: 10.1016/0005-2736(75)90101-7. [DOI] [PubMed] [Google Scholar]
  23. Weiner A. M., Platt T., Weber K. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J Biol Chem. 1972 May 25;247(10):3242–3251. [PubMed] [Google Scholar]
  24. Wilson D. L., Hall M. E., Stone G. C., Rubin R. W. Some improvements in two-dimensional gel electrophoresis of proteins. Protein mapping of eukaryotic tissue extracts. Anal Biochem. 1977 Nov;83(1):33–44. doi: 10.1016/0003-2697(77)90506-1. [DOI] [PubMed] [Google Scholar]

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