Abstract
Thylakoid membranes are laterally differentiated into appressed and non-appressed regions called grana and stroma lamellae. Pure stroma lamellae isolated from wild type maize and barley leaves contain photosystem I and its light-harvesting antennae, the cytochromeb6/f complex and coupling factor. Maize stroma lamellae contained only 2% of the total photosystem II polypeptides found in whole thylakoids, and most of the small amount of the photosystem II light-harvesting complex (LHCII) was associated with photosystem I. These results were consistent with the low rates of photosystem II electron transport and low levels of the high potential form of cytochromeb-559. Immune blot assays indicated that about half of the low potential form of cytochromeb-559 in stroma lamellae was antigenically distinct from that derived from the high potential form. The amount of LHCII in stroma lamellae could be increased by exposing leaves to bright white light (state 2) prior to the isolation of stromal lamellae. This LHCII caused a 15% increase in photosystem I antenna size and was different from the LHCII found in grana lamellae, since it lacked a 26 kD polypeptide possibly involved in thylakoid appression. These results demonstrate that the migration of LHCII from appressed to non-appressed lamellae as a result of changes in the relative amounts of energy absorbed by the 2 different photosystems, also occurs in vivo.
The reaction centre core of photosystem I was isolated from barley thylakoids and its molecular weight determined to be 650 kD. Attack by various proteases cleaved it into fragments of less than 5 kD, although the complex was still photoactive. However, the kinetics of photo-oxidation of P700 under light-limiting conditions were slower after proteolysis, indicating less efficient energy transfer. In a barley mutant lacking photosystem I, a chlorophylla species absorbing at 689 nm was lacking, accounting for about 30 molecules out of every 500 in wild type thylakoids.
Finally, a mutant completely lacking photosystem II activity,viridis −115, has been examined. It contained only 4% of the photosystem II-containing EFS particles found in wild type thylakoids, and lacked a chlorophylla species absorbing at 683 nm. Immune electron microscopy revealed that the α-subunit of cytochromeb-559 and the 33 kD polypeptide of the oxygen evolving complex were correctly located in appressed thylakoids, in spite of the lack of the major photosystem II core polypeptides. A double mutant, lacking both photosystem II and LHCII was found to contain grana, even though its thylakoids lacked the 2 complexes normally associated with maintenance of membrane appression in vivo.
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Abbreviations
- CF11/o :
-
extrinsic/intrinsic chloroplast coupling factor
- CP:
-
chlorophyll-protein
- DCMU:
-
3-(3,4-dichlorophenyl)-1,1′-dimethyl urea
- EF:
-
endoplasmic fracture face
- kD:
-
kilodaltons
- LHCI/II:
-
light-harvesting complex of PSI/II
- PF:
-
protoplasmic fracture face
- PSI/II:
-
photosystem I/II
- s:
-
stacked
- u:
-
unstacked
References
Anderson, B. &J.M. Anderson: Lateral hetero-geneity in the distribution of chlorophyll-protein complexes of the thylakoid membrane of spinach chloroplasts. Biochim. Biophys. Acta 593, 427–440 (1980)
Bassi, R., G. Giacometti &D.J. Simpson: Characterization of stroma membranes from Zea mays L. chloroplasts. Carlsberg Res. Commun. 52, 221–232 (1988a)
Bassi, R., G. Giacometti &D.J. Simpson: Changes in the organization of stroma membranes induced by in vivo state 1—state 2 transition. Biochim. Biophys. Acta 935, 165–192 (1988b)
Cox, R. &B. Anderson: Lateral and transversal organization of cytochromes in the chloroplast thylakoid membrane. Biochem. Biophys. Res. Commun. 103, 1336–1342 (1981)
Gounaris, K., A.R.R. Brain, P.J. Quinn &W.P. Williams: Structural reorganization of chloroplast thylakoid membranes in response to heat stress. Biochim. Biophys. Acta 766, 198–208 (1984)
Hinz, U.G. &D.J. Simpson: The protease-resistant core of the reaction centre protein of photosystem I. Carlsberg Res. Commun. 53, 321–330 (1988)
Hiratsuka, J., H. Shimada, R.F. Whittier, T. Ishibashi, M. Sakamoto, M. Mori, M. Kondo, Y. Honji, C.R. Sun, B.Y. Meng, Y.Q. Li, A. Kanno, Y. Nishizawa, A. Hirai, K. Shinozaki & M. Sugiura: The complete sequence of the rice (Oryza sativa) chloroplast genome. In press (1989)
Mayfield, S.P., M. Rahire, G. Frank, H. Zuber &J.-D. Rochaix: Expression of the nuclear gene encoding oxygen-evolving enhancer protein 2 is required for high levels of photosynthetic oxygen evolution inChlamydomonas reihardtii. Proc. Natl. Acad. Sci. USA 84, 749–753 (1987)
Machold, O., D.J. Simpson &G. Høyer-Hansen: Correlation between the freeze-fracture appearance and polypeptide composition of thylakoid membranes in barley. Carlsberg Res. Commun. 42, 499–516 (1977)
Ohyama, K., H. Fuzuzawa, T. Kohchi, H. Shirai, S. Sano, K. Umesono, Y. Shiki, M. Takeuchi, Z. Chang, S. Aota, H. Inukuchi &H. Ozeki: Chloroplast gene organization deduced from complete sequence of liverwortMarchantia polymorpha chloroplast DNA. Nature 322, 572–574 (1986)
Shinozaki, K., M. Ohme, M. Tanaka, T. Wakasugi, N. Hayashida, T. Matsubayashi, J. Chunwongse, J. Obokata, K. Yamaguchi-Shinozaki, C. Ohto, K. Torazawa, B.Y. Meng, M. Sugita, H. Deno, T. Kamogashira, K. Yamada, J. Kusuda, F. Takaiwa, A. Kata, N. Tohdoh, H. Shimada &M. Sugiura: The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 5, 2043–2049 (1986)
Simpson, D.J.: Freeze-fracture studies on barley plastid membranes III. Location of the light-har-vesting chlorophyll-protein. Carlsberg Res. Commun. 44, 305–336 (1979)
Simpson, D.J.: Freeze-fracture studies on barley plastid membranes VI. Location of the P700 chlorophylla-protein 1. Eur. J. Cell Biol. 31, 305–314 (1983)
Simpson, D.J.: Low temperature absorption spectroscopy of barley mutants. Gaussian deconvolution and fourth derivative analysis. Carlsberg Res. Commun. 53, 343–356 (1988)
Simpson, D.J. &D.J. Anderson: Extrinsic polypeptides of the chloroplast oxygen evolving complex constitute the tetrameric ESs particles of higher plant thylakoids. Carlsberg Res. Commun. 51, 467–474 (1986)
Simpson, D.J. &S.P. Robinson: Freeze-fracture ultrastructure of thylakoid membranes in chloroplasts from manganese-deficient plants. Plant Physiol. 74, 735–741 (1984)
Simpson, D.J., O. Vallon &D. von Wettstein: Freeze-fracture studies of barley plastid membranes VIII. Inviridis −115, a mutant completely lacking photosystem II, OEE1 and the α-subunit of cytochromeb-559 accumulate in appressed thylakoids. Biochim. Biophys. Acta 975, 164–174 (1989)
Staehelin, L.A. &C.J. Arntzen: Regulation of chloroplast membrane function: protein phosphorylation changes the spatial organization of membrane components. J. Cell Biol. 98, 1327–1337 (1983)
Tittgen, J., J. Hermans, J. Steppuhn, T. Jansen, C. Jansson, B. Anderson, R. Nechushtai, N. Nelson &R.G. Herrmann: Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins. Mol. Gen. Genet. 204, 258–265 (1986)
Vallon, O., G. Høyer-Hansen &D.J. Simpson: Photosytem II and cytochromeb-559 in the stroma lamellae of barley chloroplasts. Carlsberg Res. Commun. 52, 405–421 (1987)
Vallon, O., F.-A. Wollmann &J. Olive: Lateral distribution of the main protein complexes of the photosynthetic apparatus inChlamydomonas reinhardtii and in spinach: an immunocytochemical study using intact thylakoid membranes and a PSII-enriched membrane preparation. Photobiochem. Photobiophys. 12, 203–220 (1986)
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Simpson, D.J., Von Wettstein, D. The structure and function of the thylakoid membrane. Carlsberg Res. Commun. 54, 55 (1989). https://doi.org/10.1007/BF02907585
DOI: https://doi.org/10.1007/BF02907585