Abstract
The precursor of the major light-harvesting chlorophylla/b-proteins of photosystem II was synthesizedin vitro from a gene fromLemna gibba. When the labelled precursor was incubated with developing barley plastids, the precursor and the processed polypeptide were incorporated in the thylakoids in proportions that varied depending on the developmental stage of plastids. At early stages of development most of the precursor associated with the thylakoids could be removed by washing with 0.1 M NaOH, while in more mature plastids most of its was resistant to a NaOH wash. Insertion of the precursor into thylakoids required the presence of a stromal factor and Mg-ATP. The stromal factor is probably a protein. The insertion reaction has an optimal temperature of 25°C and a pH of 8. The appearance of the stromal factor and the thylakoid membrane's receptivity for the insertion of the precursor depended on the stage of plastid development. These observations are consistent with the hypothesis that the insertion of the precursor into the thylakoid prior to its proteolytic processing, is one of the steps involved in the assembly of the light-harvesting complex of photosystem II.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Apel K, Kloppstech K: The plastid membranes of barley (Hordeum vulgare). Light induced appearance of mRNA coding for the apoprotein of the light-harvesting chlorophylla/b protein. Eur J Biochem 85: 581–588 (1978).
Chitnis PR, Harel E, Kohorn BD, Tobin EM, Thornber JP: Assembly of the precursor and processed light-harvesting chlorophylla/b protein ofLemna into the light-harvesting complex II of barley etiochloroplasts. J Cell Biol 102: 982–988 (1986).
Chitnis PR, Nechushtai R, Harel E, Thornber JP: Some requirements for the insertion of apoproteins ofLemna light-harvesting complex II into barley thylakoids. In: Biggins J (ed.) Progress in Photosynthesis Research, Volume IV, pp 573–576. Dordrecht: Martinus Nijhoff (1987).
Chua N-H, Schmidt GW: Transport of proteins into mitochondria and chloroplasts. J Cell Biol 81: 461–483 (1979).
Cline K: Import of proteins into chloroplasts. Membrane integration of a thylakoid precursor protein reconstituted in chloroplast lysates. J Biol Chem 261: 14804–14810 (1986).
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).
Crooke E, Wickner W: The trigger factor: A soluble protein which folds pro-OmpA into a membrane assembly competent form. Proc Natl Acad Sci (USA) In press (1987).
Grossman AR, Bartlett SG, Chua N-H: Energy-dependent uptake of cytoplasmically-synthesized polypeptides by chloroplasts. Nature (Lond.) 285: 625–628 (1980).
Grossman AR, Barlett SG, Schmidt GW, Mullet JE, Chua N-H: Optimal conditions for post-translational uptake of proteins by isolated chloroplasts. J Biol Chem 257: 1558–1563 (1982).
Hiller RG, Pilger TBG, Genge S: Formation of chlorophyll protein complexes during greening of etiolated barley leaves. In: Akoyunoglou G (ed.) Chloroplast Development, pp 215–220. Elsevier/North-Holland Biomedical Press (1978).
Hoyer-Hansen G, Simpson DJ: Changes in the polypeptide composition of internal membranes of barley plastids during greening. Carlsberg Res Commun 42: 379–389 (1977).
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).
Kohorn BD, Harel E, Chitnis PR, Thornber JP, Tobin EM: Functional and mutational analysis of the light-harvesting chlorophylla/b protein of thylakoid membranes. J Cell Biol 102: 972–981 (1986).
Laemmli UK: Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature (Lond.) 227: 680–685 (1970).
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).
Mullet JE: The amino acid sequence of the polypeptide which regulates membrane adhesion (grana stacking) in chloroplasts. J Biol Chem 258: 9941–9948 (1983).
Schmidt GW, Bartlett SG, Grossman AR, Cashmore AR, Chua N-H: Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophylla/b-protein complex. J Cell Biol 91: 468–478 (1982).
Tanaka A, Tsuji H: Appearance of chlorophyll-protein complexes in greening barley seedlings. Plant Cell Physiol 26: 893–902 (1985).
Thornber JP: Biochemical characterization and structure of pigment-proteins of photosynthetic organisms. In: Staehelin LA, Arntzen CJ (eds) Encycl Plant Physiol New Series, Volume 19. Photosynthesis III: Photosynthetic membranes, pp 98–142. Berlin: Springer-Verlag (1986).
Tobin EM, Silverthorne J: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chitnis, P.R., Nechushtai, R. & Thornber, J.P. Insertion of the precursor of the light-harvesting chlorophylla/b-protein into the thylakoids requires the presence of a developmentally regulated stromal factor. Plant Mol Biol 10, 3–11 (1987). https://doi.org/10.1007/BF00014181
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00014181