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. 1984 Jul;159(1):251–259. doi: 10.1128/jb.159.1.251-259.1984

Regular surface layer of Azotobacter vinelandii.

W H Bingle, J L Doran, W J Page
PMCID: PMC215621  PMID: 6735982

Abstract

Washing Azotobacter vinelandii UW1 with Burk buffer or heating cells at 42 degrees C exposed a regular surface layer which was effectively visualized by freeze-etch electron microscopy. This layer was composed of tetragonally arranged subunits separated by a center-to-center spacing of approximately 10 nm. Cells washed with distilled water to remove an acidic major outer membrane protein with a molecular weight of 65,000 did not possess the regular surface layer. This protein, designated the S protein, specifically reattached to the surface of distilled-water-washed cells in the presence of the divalent calcium, magnesium, strontium, or beryllium cations. All of these cations except beryllium supported reassembly of the S protein into a regular tetragonal array. Although the surface localization of the S protein has been demonstrated, radioiodination of exposed envelope proteins in whole cells did not confirm this. The labeling behavior of the S protein could be explained on the basis of varying accessibilities of different tyrosine residues to iodination.

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Selected References

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

  1. Barnes E. M., Jr, Bhattacharyya P. Isolation of membrane vesicles with inverted topology by osmotic lysis of Azotobacter vinelandii spheroplasts. J Supramol Struct. 1977;6(3):333–344. doi: 10.1002/jss.400060306. [DOI] [PubMed] [Google Scholar]
  2. Bayer M. E., Leive L. Effect of ethylenediaminetetraacetate upon the surface of Escherichia coli. J Bacteriol. 1977 Jun;130(3):1364–1381. doi: 10.1128/jb.130.3.1364-1381.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beveridge T. J., Murray R. G. Dependence of the superficial layers of Spirillum putridiconchylium on Ca2+ or Sr2+. Can J Microbiol. 1976 Sep;22(9):1233–1244. doi: 10.1139/m76-183. [DOI] [PubMed] [Google Scholar]
  4. Beveridge T. J., Murray R. G. Reassembly in vitro of the superficial cell wall components of Spirillum putridiconcyhylium. J Ultrastruct Res. 1976 Apr;55(1):105–118. doi: 10.1016/s0022-5320(76)80086-x. [DOI] [PubMed] [Google Scholar]
  5. Beveridge T. J., Murray R. G. Superficial cell-wall layers on Spirillum "Ordal" and their in vitro reassembly. Can J Microbiol. 1976 Apr;22(4):567–582. doi: 10.1139/m76-085. [DOI] [PubMed] [Google Scholar]
  6. Beveridge T. J. Surface arrays on the wall of Sporosarcina ureae. J Bacteriol. 1979 Sep;139(3):1039–1048. doi: 10.1128/jb.139.3.1039-1048.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bishop P. E., Jarlenski D. M., Hetherington D. R. Evidence for an alternative nitrogen fixation system in Azotobacter vinelandii. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7342–7346. doi: 10.1073/pnas.77.12.7342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Blakesley R. W., Boezi J. A. A new staining technique for proteins in polyacrylamide gels using coomassie brilliant blue G250. Anal Biochem. 1977 Oct;82(2):580–582. doi: 10.1016/0003-2697(77)90197-x. [DOI] [PubMed] [Google Scholar]
  9. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  10. Buckmire F. L., Murray R. G. Substructure and in vitro assembly of the outer, structured layer of Spirillum serpens. J Bacteriol. 1976 Jan;125(1):290–299. doi: 10.1128/jb.125.1.290-299.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Concino M. F., Goodgal S. H. Haemophilus influenzae polypeptides involved in deoxyribonucleic acid uptake detected by cellular surface protein iodination. J Bacteriol. 1981 Oct;148(1):220–231. doi: 10.1128/jb.148.1.220-231.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DeVoe I. W., Costerton J. W., MacLeod R. A. Demonstration by freeze-etching of a single cleavage plane in the cell wall of a gram-negative bacterium. J Bacteriol. 1971 May;106(2):659–671. doi: 10.1128/jb.106.2.659-671.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Doran J. L., Page W. J. Heat sensitivity of Azotobacter vinelandii genetic transformation. J Bacteriol. 1983 Jul;155(1):159–168. doi: 10.1128/jb.155.1.159-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  15. Fisher R. J., Brill W. J. Mutants of Azotobacter vinelandii unable to fix nitrogen. Biochim Biophys Acta. 1969 Jun 17;184(1):99–105. doi: 10.1016/0304-4165(69)90103-2. [DOI] [PubMed] [Google Scholar]
  16. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  17. Harris W. F., Scriven L. E. Function of dislocations in cell walls and membranes. Nature. 1970 Nov 28;228(5274):827–829. doi: 10.1038/228827a0. [DOI] [PubMed] [Google Scholar]
  18. Irvin R. T., MacAlister T. J., Costerton J. W. Tris(hydroxymethyl)aminomethane buffer modification of Escherichia coli outer membrane permeability. J Bacteriol. 1981 Mar;145(3):1397–1403. doi: 10.1128/jb.145.3.1397-1403.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kay W. W., Buckley J. T., Ishiguro E. E., Phipps B. M., Monette J. P., Trust T. J. Purification and disposition of a surface protein associated with virulence of Aeromonas salmonicida. J Bacteriol. 1981 Sep;147(3):1077–1084. doi: 10.1128/jb.147.3.1077-1084.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Loeb M. R., Smith D. H. Lactoperoxidase and Iodo-Gen-catalyzed iodination labels inner and outer membrane proteins of Haemophilus influenzae. J Bacteriol. 1983 Jul;155(1):443–446. doi: 10.1128/jb.155.1.443-446.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  23. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  24. Moor H. Freeze-etching. Int Rev Cytol. 1969;25:391–412. doi: 10.1016/s0074-7696(08)60209-0. [DOI] [PubMed] [Google Scholar]
  25. Nakae T., Nikaido H. Outer membrane as a diffusion barrier in Salmonella typhimurium. Penetration of oligo- and polysaccharides into isolated outer membrane vesicles and cells with degraded peptidoglycan layer. J Biol Chem. 1975 Sep 25;250(18):7359–7365. [PubMed] [Google Scholar]
  26. 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]
  27. Olins A. L., Warner R. C. Physicochemical studies on a lipopolysaccharide from the cell wall of Azotobacter vinelandii. J Biol Chem. 1967 Nov 10;242(21):4994–5001. [PubMed] [Google Scholar]
  28. Page W. J., Doran J. L. Recovery of competence in calcium-limited Azotobacter vinelandii. J Bacteriol. 1981 Apr;146(1):33–40. doi: 10.1128/jb.146.1.33-40.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Page W. J., Huyer M. Derepression of the Azotobacter vinelandii siderophore system, using iron-containing minerals to limit iron repletion. J Bacteriol. 1984 May;158(2):496–502. doi: 10.1128/jb.158.2.496-502.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Page W. J., Sadoff H. L. Physiological factors affecting transformation of Azotobacter vinelandii. J Bacteriol. 1976 Mar;125(3):1080–1087. doi: 10.1128/jb.125.3.1080-1087.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Page W. J., von Tigerstrom M. Iron- and molybdenum-repressible outer membrane proteins in competent Azotobacter vinelandii. J Bacteriol. 1982 Jul;151(1):237–242. doi: 10.1128/jb.151.1.237-242.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schenk S. P., Earhart C. F. Characterization of the predominant Azotobacter vinelandii envelope protein. J Bacteriol. 1981 Apr;146(1):398–403. doi: 10.1128/jb.146.1.398-403.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schenk S. P., Earhart C. F., Wyss O. A unique envelope protein in azotobacter vinelandii. Biochem Biophys Res Commun. 1977 Aug 22;77(4):1452–1458. doi: 10.1016/s0006-291x(77)80142-3. [DOI] [PubMed] [Google Scholar]
  34. Sleytr U. B., Messner P. Crystalline surface layers on bacteria. Annu Rev Microbiol. 1983;37:311–339. doi: 10.1146/annurev.mi.37.100183.001523. [DOI] [PubMed] [Google Scholar]
  35. Sleytr U. B. Regular arrays of macromolecules on bacterial cell walls: structure, chemistry, assembly, and function. Int Rev Cytol. 1978;53:1–62. doi: 10.1016/s0074-7696(08)62240-8. [DOI] [PubMed] [Google Scholar]
  36. Thorne K. J., Thornley M. J., Naisbitt P., Glauert A. M. The nature of the attachment of a regularly arranged surface protein to the outer membrane of an Acinetobacter sp. Biochim Biophys Acta. 1975 Apr 21;389(1):97–116. doi: 10.1016/0005-2736(75)90388-0. [DOI] [PubMed] [Google Scholar]
  37. Thornley M. J., Thorne K. J., Glauert A. M. Detachment and chemical characterization of the regularly arranged subunits from the surface of an Acinetobacter. J Bacteriol. 1974 May;118(2):654–662. doi: 10.1128/jb.118.2.654-662.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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