Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Oct;168(1):167–172. doi: 10.1128/jb.168.1.167-172.1986

Effect of uncoupler on assembly pathway for pigment-binding protein of bacterial photosynthetic membranes.

R Dierstein, G Drews
PMCID: PMC213433  PMID: 3531166

Abstract

The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) was used to investigate membrane protein assembly in the phototrophic bacterium Rhodobacter capsulatus. As found for Escherichia coli (T. Date, G. Zwizinsky, S. Ludmerer, and W. Wickner, Proc. Natl. Acad. Sci. 77:827-831, 1980) and mitochondrial proteins (N. Nelson and G. Schatz, Proc. Natl. Acad. Sci. USA 76:4365-4369, 1979), assembly across the bacterial photosynthetic membranes was sensitive to CCCP. At uncoupler concentrations which were sufficient to block the export of the periplasmic cytochrome c2 and an outer membrane protein, the integration of pigment-binding protein into the photosynthetic apparatus was abolished. The unassembled protein was detected on the inner surface of the intracytoplasmic membrane. After inactivation of CCCP, accumulated protein continued insertion into the membrane. The data suggest that after binding to the cytoplasmic face of the membrane, translocation of protein into a transmembrane orientation takes place, which is a prerequisite for the formation of a functional pigment-protein complex.

Full text

PDF
167

Images in this article

168Image
on p.168
169Image
on p.169
169Image
on p.169
170Image
on p.170

Selected References

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

  1. Anderson D. J., Mostov K. E., Blobel G. Mechanisms of integration of de novo-synthesized polypeptides into membranes: signal-recognition particle is required for integration into microsomal membranes of calcium ATPase and of lens MP26 but not of cytochrome b5. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7249–7253. doi: 10.1073/pnas.80.23.7249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Broglie R. M., Hunter C. N., Delepelaire P., Niederman R. A., Chua N. H., Clayton R. K. Isolation and characterization of the pigment-protein complexes of Rhodopseudomonas sphaeroides by lithium dodecyl sulfate/polyacrylamide gel electrophoresis. Proc Natl Acad Sci U S A. 1980 Jan;77(1):87–91. doi: 10.1073/pnas.77.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brusilow W. S., Gunsalus R. P., Hardeman E. C., Decker K. P., Simoni R. D. In vitro synthesis of the F0 and F1 components of the proton translocating ATPase of Escherichia coli. J Biol Chem. 1981 Apr 10;256(7):3141–3144. [PubMed] [Google Scholar]
  4. Bélanger G., Bérard J., Gingras G. Isolation and partial characterization of the messenger RNA encoding the B880 holochrome protein of Rhodospirillum rubrum. Eur J Biochem. 1985 Dec 16;153(3):477–484. doi: 10.1111/j.1432-1033.1985.tb09326.x. [DOI] [PubMed] [Google Scholar]
  5. Chung C. H., Goldberg A. L. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931–4935. doi: 10.1073/pnas.78.8.4931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daniels C. J., Bole D. G., Quay S. C., Oxender D. L. Role for membrane potential in the secretion of protein into the periplasm of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5396–5400. doi: 10.1073/pnas.78.9.5396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Date T., Zwizinski C., Ludmerer S., Wickner W. Mechanisms of membrane assembly: effects of energy poisons on the conversion of soluble M13 coliphage procoat to membrane-bound coat protein. Proc Natl Acad Sci U S A. 1980 Feb;77(2):827–831. doi: 10.1073/pnas.77.2.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dierstein R., Drews G. Membrane differentiation and assembly of the pigment-protein complexes of the photosynthetic bacterium Rhodopseudomonas capsulata. Prog Clin Biol Res. 1982;102(Pt B):247–256. [PubMed] [Google Scholar]
  9. Dierstein R. Synthesis of pigment-binding protein in toluene-treated Rhodopseudomonas capsulata and in cell-free systems. Eur J Biochem. 1984 Feb 1;138(3):509–518. doi: 10.1111/j.1432-1033.1984.tb07945.x. [DOI] [PubMed] [Google Scholar]
  10. Dierstein R., Wickner W. The leader region of pre-maltose binding protein binds amphiphiles. A model for self-assembly in protein export. J Biol Chem. 1985 Dec 15;260(29):15919–15924. [PubMed] [Google Scholar]
  11. Drews G. Structure and functional organization of light-harvesting complexes and photochemical reaction centers in membranes of phototrophic bacteria. Microbiol Rev. 1985 Mar;49(1):59–70. doi: 10.1128/mr.49.1.59-70.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ebeling W., Hennrich N., Klockow M., Metz H., Orth H. D., Lang H. Proteinase K from Tritirachium album Limber. Eur J Biochem. 1974 Aug 15;47(1):91–97. doi: 10.1111/j.1432-1033.1974.tb03671.x. [DOI] [PubMed] [Google Scholar]
  13. Enequist H. G., Hirst T. R., Harayama S., Hardy S. J., Randall L. L. Energy is required for maturation of exported proteins in Escherichia coli. Eur J Biochem. 1981 May 15;116(2):227–233. doi: 10.1111/j.1432-1033.1981.tb05323.x. [DOI] [PubMed] [Google Scholar]
  14. Engelman D. M., Steitz T. A. The spontaneous insertion of proteins into and across membranes: the helical hairpin hypothesis. Cell. 1981 Feb;23(2):411–422. doi: 10.1016/0092-8674(81)90136-7. [DOI] [PubMed] [Google Scholar]
  15. Hochman A., Fridberg I., Carmeli C. The location and function of cytochrome c2 in Rhodopseudomonas capsulate membranes. Eur J Biochem. 1975 Oct 1;58(1):65–72. doi: 10.1111/j.1432-1033.1975.tb02349.x. [DOI] [PubMed] [Google Scholar]
  16. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Meister H., Bachofen R., Semenza G., Brunner J. Membrane topology of light-harvesting protein B870-alpha of Rhodospirillum rubrum G-9+. Amino acid residues in contact with the lipid bilayer as inferred from labeling with photogenerated carbenes. J Biol Chem. 1985 Dec 25;260(30):16326–16331. [PubMed] [Google Scholar]
  19. Nelson N., Schatz G. Energy-dependent processing of cytoplasmically made precursors to mitochondrial proteins. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4365–4369. doi: 10.1073/pnas.76.9.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Randall L. L., Hardy S. J. Export of protein in bacteria. Microbiol Rev. 1984 Dec;48(4):290–298. doi: 10.1128/mr.48.4.290-298.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schmidt G. W., Bartlett S. G., Grossman A. R., Cashmore A. R., Chua N. H. Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol. 1981 Nov;91(2 Pt 1):468–478. doi: 10.1083/jcb.91.2.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tadros M. H., Frank R., Drews G. Localization of the exposed N-terminal region of the B800-850 alpha and beta light-harvesting polypeptides on the cytoplasmic surface of Rhodopseudomonas capsulata chromatophores. J Bacteriol. 1986 Jul;167(1):96–100. doi: 10.1128/jb.167.1.96-100.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tadros M. H., Suter F., Drews G., Zuber H. The complete amino-acid sequence of the large bacteriochlorophyll-binding polypeptide from light-harvesting complex II (B800-850) of Rhodopseudomonas capsulata. Eur J Biochem. 1983 Jan 1;129(3):533–536. doi: 10.1111/j.1432-1033.1983.tb07081.x. [DOI] [PubMed] [Google Scholar]
  24. Weckesser J., Zalman L. S., Nikaido H. Porin from Rhodopseudomonas sphaeroides. J Bacteriol. 1984 Jul;159(1):199–205. doi: 10.1128/jb.159.1.199-205.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wickner W. T., Lodish H. F. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. doi: 10.1126/science.4048938. [DOI] [PubMed] [Google Scholar]
  26. Wolfe P. B., Wickner W. Bacterial leader peptidase, a membrane protein without a leader peptide, uses the same export pathway as pre-secretory proteins. Cell. 1984 Apr;36(4):1067–1072. doi: 10.1016/0092-8674(84)90056-4. [DOI] [PubMed] [Google Scholar]
  27. Youvan D. C., Bylina E. J., Alberti M., Begusch H., Hearst J. E. Nucleotide and deduced polypeptide sequences of the photosynthetic reaction-center, B870 antenna, and flanking polypeptides from R. capsulata. Cell. 1984 Jul;37(3):949–957. doi: 10.1016/0092-8674(84)90429-x. [DOI] [PubMed] [Google Scholar]
  28. Youvan D. C., Ismail S. Light-harvesting II (B800-B850 complex) structural genes from Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1985 Jan;82(1):58–62. doi: 10.1073/pnas.82.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zimmermann R., Watts C., Wickner W. The biosynthesis of membrane-bound M13 coat protein. Energetics and assembly intermediates. J Biol Chem. 1982 Jun 10;257(11):6529–6536. [PubMed] [Google Scholar]
  30. von Heijne G., Blomberg C. Trans-membrane translocation of proteins. The direct transfer model. Eur J Biochem. 1979 Jun;97(1):175–181. doi: 10.1111/j.1432-1033.1979.tb13100.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES