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The apoprotein precursor of the major light-harvesting complex of photosystem II (LHCIIb) is inserted primarily into stromal lamellae and subsequently migrates to the grana

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Abstract

The formation of the lateral distribution of the major antenna complex of photosystem II (LHCIIb) between the granal and stromal lamellae was studied. Specifically, the localization of the insertion and the assembly of the precursor of the apoprotein of LHCIIb (pLHCP) were studied with isolated thylakoids. After insertion of pLHCP into isolated thylakoids, fractionation of the latter into granal and stromal lamellar was performed. At 25 °C most of the precursor was located in the granal lamellae, although both highly purified granal and stromal lamellar fractions demonstrated a similar capability to insert pLHCP. When the insertion reaction to the thylakoids was performed at 10 °C, followed by their separation into stromal and granal lamellae, the labelled pLHCP was localized in the stromal ones. To examine whether pLHCP inserts into both granal and stromal lamellae, or preferentially into stromal lamellae and subsequently migrating to granal lamellae, a chase experiment was performed. Insertion of pLHCP at 10 °C was followed by chase of the radioactive precursor with excess of non-radioactive pLHCP at 25 °C. From the results presented it is evident that the level of pLHCP in stromal lamellae was gradually reduced, while it gradually accumulated in the granal lamellae. Furthermore, the pLHCP in the stromal lamellae was found to be in a ‘free’ form, while after migrating to the granal lamellae it assembled into the pigmented LHCIIb.

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References

  1. Thornber JP: Biochemical characterization and structure of pigment-proteins of photosynthetic organisms. In: Staehelin LA, Arntzen CJ (ed) Encyclopedia of Plant Physiology, New Series, vol 19. Photosynthesis III: Photosynthetic membranes, pp. 98–142. Springer-Verlag, Berlin (1986).

    Google Scholar 

  2. Thornber JP, Peter GF, Nechushtai R, Chitnis PR, Hunter FA, Tobin EM: Electrophoretic separation of chlorophyll-protein complexes and their apoproteins. In: Akoyonoglu G (ed) Regulation of Chloroplast Development, pp. 249–258. Alan R. Liss New York (1986).

    Google Scholar 

  3. Bennet J: Regulation of photosynthesis by reversible phosphorylation of the light-harvesting chlorophyll a/b protein. Biochem J 212: 1–13 (1983).

    PubMed  Google Scholar 

  4. Machold O, Meister A: Resolution of the light-harvesting chlorophyll a/b-protein complexes. Biochim Biophys Acta 546: 472–480 (1979).

    PubMed  Google Scholar 

  5. Leutwiler LS, Meyerowitz EM, Tobin EM: Structure and expression of three light-harvesting chlorophyll a/b-binding protein genes in Arabidopsis thaliana. Nucleic Acids Res 14: 4051–4064 (1986).

    PubMed  Google Scholar 

  6. Dunsmuir P, Bedbrook J: Chlorophyll a/b-binding proteins and the small subunit of ribulose biphosphate carboxylase are encoded by multiple genes in petunia. NATO-ASI Series A, pp. 63–221 (1983).

  7. Dunsmuir P, Smith SM, Bedbrook J: The major chlorophyll a/b-binding protein of petunia is composed of several polypeptides encoded by a number of distinct nuclear genes. J Mol Appl Genet 2: 285–300 (1983).

    PubMed  Google Scholar 

  8. Chitnis PR, Thornber JP: The major light-harvesting complex of photosystem-II. Aspects of its molecular and cell biology. Photosynthesis Res 16: 41–63 (1988).

    Google Scholar 

  9. Grossman A, Bartlett S, Chua N-H: Energy-dependent uptake of cytoplasmically synthesized polypeptides by chloroplasts. Nature 285: 625–628 (1980).

    Google Scholar 

  10. Schmidt GW, Bartlett SG, Grossman AR, Cashmore AR, Chua N-H: Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol 91: 468–478 (1981).

    Article  PubMed  Google Scholar 

  11. Chitnis PR, Harel E, Kohorn BD, Tobin EM, Thornber JP: Assembly of the precursor and processed light-harvesting chlorophyll a/b protein of Lemna into the light-harvesting complex II of barley etiochloroplasts. J Cell Biol 102: 982–988 (1986).

    Article  PubMed  Google Scholar 

  12. Cline K, Werner-Washburne M, Lubben TH, Keegstra K: Precursors to two nuclear-encoded chloroplast proteins bind to the outer envelope membrane before being imported into chloroplasts. J Biol Chem 260: 3691–3696 (1985).

    PubMed  Google Scholar 

  13. Kohorn BD, Harel E, Chitnis PR, Thornber JP, Tobin EM: Functional and mutational analysis of the light-harvesting chlorophyll a/b protein of thylakoid membranes. J Cell Biol 102: 972–981 (1986).

    Article  PubMed  Google Scholar 

  14. Chitnis PR, Morishige DT, Nechushtai R, Thornber JP: Assembly of the barley light-harvesting chlorophyll a/b proteins in barley etiochloroplasts involves processing of the precursor on thylakoids. Plant Mol Biol 11: 95–107 (1988).

    Google Scholar 

  15. Chitnis PR, Nechushtai R, Harel E, Thornber JP: Some requirement for the insertion of the precursor of apoproteins of Lemna light-harvesting complex II into barley thylakoids. In: Biggins J (ed) Progress in Photosynthesis Research, volume IV, pp. 573–576. Martinus Nijhoff/Junk. Dordrecht (1987).

    Google Scholar 

  16. Chitnis PR, Nechushtai R, Thornber JP: Insertion of the precursor of the light-harvesting chlorophyll a/b-protein into thylakoids requires the presence of a developmentally regulated stromal factor. Plant Mol Biol 10: 3–11 (1987).

    Google Scholar 

  17. Cline K: Import of proteins into chloroplasts: membrane integration of thylakoid precursor protein reconstituted in chloroplast lysates. J Biol Chem 261: 14804–14810 (1986).

    PubMed  Google Scholar 

  18. Cline K: Light-harvesting chlorophyll a/b protein: membrane insertion, proteolytic processing, assembly into LHC II and localization to appressed membranes occurs in chloroplast lysates. Plant Physiol 86: 1120–1126 (1988).

    Google Scholar 

  19. Fulsom DR, Cline K: A soluble protein factor is required in vitro for membrane insertion of the thylakoid precursor protein, pLHCP. Plant Physiol 88: 1146–1153 (1988).

    Google Scholar 

  20. Andersson B, Anderson JM: The chloroplast thylakoid membrane, isolation subfractionation and purification of its supramolecular complexes. In: Linskens HF, Jackson JF (ed) Modern Methods of Plant Analysis, New Series, vol I, pp. 231–258. Springer-Verlag, Berlin (1985).

    Google Scholar 

  21. Goodchild DJ, Highkin HE, Boardman NK: The fine structure of chloroplasts in a barley mutant lacking chlorophyll b. Exp Cell Res 43: 684–688 (1966).

    PubMed  Google Scholar 

  22. Nielsen NC, Smillie RM, Henningsen KW, vonWettstein D: Composition and function of thylakoid membranes from grana rich and grana deficient chloroplast mutants of barley. Plant Physiol 63: 174–182 (1979).

    Google Scholar 

  23. Schuster G, Ohad I, Martineau B, Taylor WC: Differentiation and development of bundle sheath and mesophyl thylakoids in maize. J Biol Chem 260: 11866–11873 (1985).

    PubMed  Google Scholar 

  24. Armond PA, Arntzen CJ, Briantais J, Vermotte C: Differentiation of chloroplast lamellae light-harvesting efficiency and grana development. Arch Biochem Biophys 175: 54–63 (1976).

    PubMed  Google Scholar 

  25. Mullet JE, Arntzen CJ: Simulation of grana stacking in a model membrane system mediated by a purified light-harvesting pigment-protein complex from chloroplast. Biochim Biophys Acta 589: 100–117 (1980).

    PubMed  Google Scholar 

  26. Ryrie IJ, Anderson JM, Goodchild DJ: The role of light harvesting chlorophyll a/b protein complex in chloroplast membrane stacking. Cation-induced aggregation of reconstituted proteoliposomes. Eur J Biochem 107: 345–354 (1980).

    PubMed  Google Scholar 

  27. Vainstein A, Ferreira P, Peterson CC, Verbeke JA, Thornber JP: Expression of the major light-harvesting chlorophyll a/b-protein and its import into thylakoids of mesophyll and bundle-sheath chloroplasts of maize. Plant Physiol 89: 602–609 (1989).

    Google Scholar 

  28. Leto KJ, Bell E, McIntosh L: Nuclear mutation leads to an accelerated turnover of chloroplast-encoded 48 kd and 34.5 kd polypeptides in thylakoids lacking photosystem II. EMBO J 4: 1645–1653 (1985).

    Google Scholar 

  29. Laemmli UK: Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).

    PubMed  Google Scholar 

  30. Peter GF, Thornber JP: Electrophoretic procedures for fractionation of photosystem I and photosystem II pigment protein of higher plants and for determination of their subunits composition. In: Rogers LJ (ed) Methods in Plant Biochemistry, vol 2: Amino Acids, Proteins, and Nucleic Acids (1989) (in press).

  31. Kamienietzky A, Nelson N: Preparation and properties of chloroplasts depleted of chloroplast coupling factor 1 by sodium bromide treatment. Plant Physiol 55: 282–287 (1975).

    Google Scholar 

  32. Matoo AK, Edelman M: Intramembrane translocation and posttranslational palmitoylation of the chloroplast 32-kDa herbicide-binding protein. Proc Natl Acad Sci USA 84: 1497–1501 (1987).

    PubMed  Google Scholar 

  33. Ellis J: Proteins as molecular chaperons. Nature 328: 378–379 (1987).

    PubMed  Google Scholar 

  34. Barber J: Influence of surface charges on thylakoid structure and function. Ann Rev Plant Physiol 33: 261–295 (1982).

    Google Scholar 

  35. Lilley RHC, Fitzgerald MP, Rieutis KG, Walker DA: Criteria of intactness and the photosynthetic activity of spinach chloroplast preparation. New Phytol 75: 1–10 (1975).

    Google Scholar 

  36. Kirwin PM, Elderfield PD, Williams RS, Robinson C: Transport of proteins into chloroplast. Organization, orientation, and lateral distribution of the plastocyanine processing peptidase in the thylakoid network. J Biol Chem 263: 18128–18132 (1988).

    PubMed  Google Scholar 

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Yalovsky, S., Schuster, G. & Nechushtai, R. The apoprotein precursor of the major light-harvesting complex of photosystem II (LHCIIb) is inserted primarily into stromal lamellae and subsequently migrates to the grana. Plant Mol Biol 14, 753–764 (1990). https://doi.org/10.1007/BF00016508

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  • DOI: https://doi.org/10.1007/BF00016508

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