Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Mar 1;102(3):982–988. doi: 10.1083/jcb.102.3.982

Assembly of the precursor and processed light-harvesting chlorophyll a/b protein of Lemna into the light-harvesting complex II of barley etiochloroplasts

PMCID: PMC2114146  PMID: 3512584

Abstract

When the in vitro synthesized precursor of a light-harvesting chlorophyll a/b binding protein (LHCP) from Lemna gibba is imported into barley etiochloroplasts, it is processed to a single form. Both the processed form and the precursor are found in the thylakoid membranes, assembled into the light-harvesting complex of photosystem II. Neither form can be detected in the stromal fraction. The relative amounts of precursor and processed forms observed in the thylakoids are dependent on the developmental stage of the plastids used for uptake. The precursor as well as the processed form can also be detected in thylakoids of greening maize plastids used in similar uptake experiments. This detection of a precursor in the thylakoids, which has not been previously reported, could be a result of using rapidly developing plastids and/or using an heterologous system. Our results demonstrate that the extent of processing of LHCP precursor is not a prerequisite for its inclusion in the complex. They are also consistent with the possibility that the processing step can occur after insertion of the protein into the thylakoid membrane.

Full Text

The Full Text of this article is available as a PDF (850.3 KB).

Selected References

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

  1. Andersson B., Anderson J. M., Ryrie I. J. Transbilayer organization of the chlorophyll-proteins of spinach thylakoids. Eur J Biochem. 1982 Apr 1;123(2):465–472. doi: 10.1111/j.1432-1033.1982.tb19790.x. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bennett J. Biosynthesis of the light-harvesting chlorophyll a/b protein. Polypeptide turnover in darkness. Eur J Biochem. 1981 Aug;118(1):61–70. doi: 10.1111/j.1432-1033.1981.tb05486.x. [DOI] [PubMed] [Google Scholar]
  4. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Chua N. H., Schmidt G. W. Transport of proteins into mitochondria and chloroplasts. J Cell Biol. 1979 Jun;81(3):461–483. doi: 10.1083/jcb.81.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cline K., Andrews J., Mersey B., Newcomb E. H., Keegstra K. Separation and characterization of inner and outer envelope membranes of pea chloroplasts. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3595–3599. doi: 10.1073/pnas.78.6.3595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cline K., Werner-Washburne M., Andrews J., Keegstra K. Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts. Plant Physiol. 1984 Jul;75(3):675–678. doi: 10.1104/pp.75.3.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Grebanier A. E., Coen D. M., Rich A., Bogorad L. Membrane proteins synthesized but not processed by isolated maize chloroplasts. J Cell Biol. 1978 Sep;78(3):734–746. doi: 10.1083/jcb.78.3.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. Markwell J. P., Thornber J. P., Boggs R. T. Higher plant chloroplasts: Evidence that all the chlorophyll exists as chlorophyll-protein complexes. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1233–1235. doi: 10.1073/pnas.76.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mishkind M. L., Wessler S. R., Schmidt G. W. Functional determinants in transit sequences: import and partial maturation by vascular plant chloroplasts of the ribulose-1,5-bisphosphate carboxylase small subunit of Chlamydomonas. J Cell Biol. 1985 Jan;100(1):226–234. doi: 10.1083/jcb.100.1.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mullet J. E. The amino acid sequence of the polypeptide segment which regulates membrane adhesion (grana stacking) in chloroplasts. J Biol Chem. 1983 Aug 25;258(16):9941–9948. [PubMed] [Google Scholar]
  17. Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Robinson C., Ellis R. J. Transport of proteins into chloroplasts. Partial purification of a chloroplast protease involved in the processing of important precursor polypeptides. Eur J Biochem. 1984 Jul 16;142(2):337–342. doi: 10.1111/j.1432-1033.1984.tb08291.x. [DOI] [PubMed] [Google Scholar]
  19. Robinson C., Ellis R. J. Transport of proteins into chloroplasts. The precursor of small subunit of ribulose bisphosphate carboxylase is processed to the mature size in two steps. Eur J Biochem. 1984 Jul 16;142(2):343–346. doi: 10.1111/j.1432-1033.1984.tb08292.x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Wolosiuk R. A., Buchanan B. B. Studies on the regulation of chloroplast NADP-linked glyceraldehyde-3-phosphate dehydrogenase. J Biol Chem. 1976 Oct 25;251(20):6456–6461. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES