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. 1989 Jan;1(1):159–166. doi: 10.1105/tpc.1.1.159

A hydrophobic, carboxy-proximal region of a light-harvesting chlorophyll a/b protein is necessary for stable integration into thylakoid membranes.

B D Kohorn 1, E M Tobin 1
PMCID: PMC159747  PMID: 2535463

Abstract

Proteins synthesized as soluble precursors in the cytoplasm of eukaryotic cells often cross organellar membrane barriers and then insert into lipid bilayers. One such polypeptide, the light-harvesting chlorophyll a/b-binding protein (LHCP), must also associate with pigment molecules and be assembled into the photosystem II light-harvesting complex in the chloroplast thylakoid membrane. A study of the import of mutant LHCPs into isolated chloroplasts has shown that a putative alpha-helical membrane-spanning domain near the carboxy terminus (helix 3) is essential for the stable insertion of LHCP in the thylakoid. Protease digestion experiments are consistent with the carboxy terminus of the protein being in the lumen. This report also shows that helix 3, when fused to a soluble protein, can target it to the thylakoids of isolated, intact chloroplasts. Although helix 3 is required for the insertion of LHCP and mutant derivatives into the thylakoid, the full insertion of helix 3 itself requires additionally the presence of other regions of LHCP. Thus, LHCP targeting and integration into thylakoid membranes requires a complex interaction involving a number of different domains of the LHCP polypeptide.

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Selected References

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

  1. Bellemare G., Bartlett S. G., Chua N. H. Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the chlorina f2 mutant of barley. J Biol Chem. 1982 Jul 10;257(13):7762–7767. [PubMed] [Google Scholar]
  2. Cline K. Import of proteins into chloroplasts. Membrane integration of a thylakoid precursor protein reconstituted in chloroplast lysates. J Biol Chem. 1986 Nov 5;261(31):14804–14810. [PubMed] [Google Scholar]
  3. Cline K., Werner-Washburne M., Lubben T. H., Keegstra K. Precursors to two nuclear-encoded chloroplast proteins bind to the outer envelope membrane before being imported into chloroplasts. J Biol Chem. 1985 Mar 25;260(6):3691–3696. [PubMed] [Google Scholar]
  4. Dalbey R. E., Wickner W. Characterization of the internal signal-anchor domain of Escherichia coli leader peptidase. J Biol Chem. 1988 Jan 5;263(1):404–408. [PubMed] [Google Scholar]
  5. Eisenberg D. Three-dimensional structure of membrane and surface proteins. Annu Rev Biochem. 1984;53:595–623. doi: 10.1146/annurev.bi.53.070184.003115. [DOI] [PubMed] [Google Scholar]
  6. Grossman A. R., Bartlett S. G., Schmidt G. W., Mullet J. E., Chua N. H. Optimal conditions for post-translational uptake of proteins by isolated chloroplasts. In vitro synthesis and transport of plastocyanin, ferredoxin-NADP+ oxidoreductase, and fructose-1,6-bisphosphatase. J Biol Chem. 1982 Feb 10;257(3):1558–1563. [PubMed] [Google Scholar]
  7. Karlin-Neumann G. A., Kohorn B. D., Thornber J. P., Tobin E. M. A chlorophyll a/b-protein encoded by a gene containing an intron with characteristics of a transposable element. J Mol Appl Genet. 1985;3(1):45–61. [PubMed] [Google Scholar]
  8. Keegstra K., Bauerle C. Targeting of proteins into chloroplasts. Bioessays. 1988 Jul;9(1):15–19. doi: 10.1002/bies.950090105. [DOI] [PubMed] [Google Scholar]
  9. Kohorn B. D., Harel E., Chitnis P. R., Thornber J. P., Tobin E. M. Functional and mutational analysis of the light-harvesting chlorophyll a/b protein of thylakoid membranes. J Cell Biol. 1986 Mar;102(3):972–981. doi: 10.1083/jcb.102.3.972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kohorn B. D., Tobin E. M. Amino acid charge distribution influences the assembly of apoprotein into light-harvesting complex II. J Biol Chem. 1987 Sep 25;262(27):12897–12899. [PubMed] [Google Scholar]
  11. Kowalski D., Kroeker W. D., Laskowski M., Sr Mung bean nuclease I. Physical, chemical, and catalytic properties. Biochemistry. 1976 Oct 5;15(20):4457–4463. doi: 10.1021/bi00665a019. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  14. Li J. Light-harvesting chlorophyll a/b-protein: Three-dimensional structure of a reconstituted membrane lattice in negative stain. Proc Natl Acad Sci U S A. 1985 Jan;82(2):386–390. doi: 10.1073/pnas.82.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McCrea P. D., Engelman D. M., Popot J. L. Topography of integral membrane proteins: hydrophobicity analysis vs. immunolocalization. Trends Biochem Sci. 1988 Aug;13(8):289–290. doi: 10.1016/0968-0004(88)90120-x. [DOI] [PubMed] [Google Scholar]
  16. Pfanner N., Hartl F. U., Neupert W. Import of proteins into mitochondria: a multi-step process. Eur J Biochem. 1988 Aug 1;175(2):205–212. doi: 10.1111/j.1432-1033.1988.tb14185.x. [DOI] [PubMed] [Google Scholar]
  17. Plumley F. G., Schmidt G. W. Reconstitution of chlorophyll a/b light-harvesting complexes: Xanthophyll-dependent assembly and energy transfer. Proc Natl Acad Sci U S A. 1987 Jan;84(1):146–150. doi: 10.1073/pnas.84.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schmidt G. W., Mishkind M. L. The transport of proteins into chloroplasts. Annu Rev Biochem. 1986;55:879–912. doi: 10.1146/annurev.bi.55.070186.004311. [DOI] [PubMed] [Google Scholar]
  19. Smeekens S., Bauerle C., Hageman J., Keegstra K., Weisbeek P. The role of the transit peptide in the routing of precursors toward different chloroplast compartments. Cell. 1986 Aug 1;46(3):365–375. doi: 10.1016/0092-8674(86)90657-4. [DOI] [PubMed] [Google Scholar]
  20. Steinback K. E., Burke J. J., Arntzen C. J. Evidence for the role of surface-exposed segments of the light-harvesting complex in cation-mediated control of chloroplast structure and function. Arch Biochem Biophys. 1979 Jul;195(2):546–557. doi: 10.1016/0003-9861(79)90381-3. [DOI] [PubMed] [Google Scholar]
  21. Verner K., Schatz G. Protein translocation across membranes. Science. 1988 Sep 9;241(4871):1307–1313. doi: 10.1126/science.2842866. [DOI] [PubMed] [Google Scholar]

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