Abstract
In the present study, using specific antibody against D1 protein, we detected four aggregates of D1 protein in thylakoid membranes from spinach leaves illuminated at high light (800–2500 μmol photons·m−2·s−1) for 3 h. Their accumulations were dependent on the light intensity to which the leaves had been subjected. Further immunoblot analysis indicated that 70 kD aggregate was a product of D1 protein cross-linked with CP43, 65 and 60 kD aggregate were two cross-linked products between D1 and D2 proteins, and 41 kD aggregate was one cross-linked D1 with α-subunit of cytochrome b 559 (Cyt b 559). This result provided the evidence for the existence of the aggregation of the D1 protein in vivo. The maximal level of D1/Cyt b 559 aggregate occurred at 1000 μmol photons·m−2·s−1 but drastically decreased with further increasing light intensity. Immunoblot analysis with phosphothreonine (Thr (P)) antibody indicated that D1/CP43 and D1/Cyt b 559 aggregates contained the phosphorylated protein(s). In vitro dephosphorylation experiment also showed that D1/Cyt b 559 and D1/CP43 aggregates lost the immunoreactivity with Thr (P) antibody after the phosphatase treatment of the membranes from high-light-illuminated leaves. Our results demonstrated that strong illumination of spinach leaves induced cross-linked products of D1 protein with its nearby polypeptides of PS, some of which co.n-tained the phosphorylated D1 protein.
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Barber, J., Nield, J., Morris, E. P. et al., The structure, function and dynamics of photosystem two, Physiol. Plant, 1997, 100(4): 817–827.
Aro, E. M., Virgin, I., Andersson, B., Photoinhibition of photosystem II. Inactivation, protein damage and turnover, Biochim. Bio-phys. Acta, 1993, 1143(2): 113–134.
Aro, E. M., Photodamage and D1 protein turnover in photosystem II, in Regulation of Photosynthesis (eds. Aro, E. M., Andersson, B.), Dordrecht: Kluwer, 2001, 377–393.
Zhang, L. X. Aro, E. M., Synthesis, membrane insertion and assembly of the chloroplast-encoded D1 protein into photosystem II, FEBS Lett., 2002, 512(1-3): 13–18.
Mattoo, A. K., Hoffman-Falk, H., Marder, J. B. et al., Regulation of protein metabolism: Coupling of photosynthetic electron transport to in vivo degradation of the rapidly metabolized 32-kilodalton protein of the chloroplast membranes, Proc. Natl. Acad. Sci. USA, 2001, 81(5): 1380–1384.
Rintamäki, E., Salonen, M., Suoranta, U. M. et al., Phosphorylation of light-harvesting complex and photosystem II core proteins shows different irradiance-dependent regulation in vivo. Application of phosphothreonine antibodies to analysis of thylakoid phosphoproteins, J. Biol. Chem., 1997, 272(48): 30476–30482.
Pursiheimo, S., Martinsuo, E., Rintamäki, E. et al., Photosystem protein phosphorylation follows four distinctly different regulatory patterns induced by environmental cues, Plant Cell Environ., 2003, 26(12): 1995–2003.
Andersson, B., Aro, E. M., Proteolytic activities and proteases of plant chloroplasts, Physiol. Plant, 1997, 100(4): 780–793.
Rintamäki, E., Kettunen, R., Aro, E. M., Differential D1 dephosphorylation in functional and photodamaged photosystem II centers, J. Biol. Chem., 1996, 271(25): 14870–14875.
Koivuniemi, A., Aro, E. M., Andersson, B., Degradation of the D1-and D2-proteins of photosystem II in higher plants is regulated by reversible phosphorylation, Biochemistry, 1995, 34(49): 16022–16029.
Barbato, R., Friso, G., Rigoni, F. et al., Characterization of a 41 kD photoinhibition adduct in isolated photosystem II reaction centers, FEBS Lett., 1992, 309(2): 165–169.
Barbato, R., Friso, G., Ponticos, M. et al., Characterization of the light-induced cross-linking of the α-subunit of cytochrome b 559 and the D1 protein in isolated photosystem II reaction centers, J. Biol. Chem., 1995, 270(41): 24032–24037.
Yamamoto, Y., Akasaka, T., Degradation of antenna chlorophyll-binding protein CP43 during photoinhibition of photosystem II, Biochemistry, 1995, 43(28): 9038–9045.
Ishikawa, Y., Nakatani, E., Henmi, T. et al., Turnover of the aggregates and cross-linked products of the D1 protein generated by acceptor-side photoinhibition of photosystem II, Biochim. Biophys. Acta, 1999, 1413(3): 147–158.
Henmi, T., Yamasaki, H., Sakuma, S. et al., Dynamic interaction between the D1 Protein, CP43 and OEC33 at the lumenal side of photosystem II in spinach chloroplasts: Evidence from light-induced cross-linking of the proteins in the donor-side photoinhibition, Plant Cell Physiol., 2003, 44(4): 451–456.
Yamamoto, Y., Quality control of photosystem II, Plant Cell Physiol., 2001, 42(2): 121–128.
Arnon, D. I., Copper enzymes in isolated chloroplasts polyphenoloxidase in Beta vulgaris, Plant Physiol., 1949, 24(1): 1–15.
Liu, Y. Q., Du, L. F., Purification and partial characterization of a phosphatase on thylakoid membrane in Ipomoea aquatica, Chin. J. Appl. Environ. Biol. (in Chinese), 2003, 9(3): 239–242.
Du, L. F., Tang, X. S., Liang, H. G., Influence of calcium ion on photosystem II oxygen evolution, Science in China, Ser. B, 1995, 38 (12): 1439-1447.
Li, X. P., Du, L. F., Ling, H. G. et al., Preparation and identification of anti-dodecapeptide of polypeptide D1 of photosystem II reaction center, Prog. Biochem. Biophys. (in Chinese), 1997, 24(3): 283–285.
Li, S. C., Du, L. F., Isolation of Mr33000 manganese-stabilizing protein and preparation of antiserum, Plant Physiol. Commu. (in Chinese), 1996, 32(6): 439–441.
Li, J., Du, L. F., A new approach to detect plant thykaloid phosphoprotein in vivo, Prog. Biochem. Biophys. (in Chinese), 2001, 28(5): 740–743.
Kettunen, R., Tyystjärvi, E., Aro, E. M., Degradation pattern of photosystem II reaction center protein D1 in intact leaves, Plant Physiol., 1996, 111(4): 1183–1190.
Rintamäki, E., Aro, E. M., Phosphorylation of photosystem II proteins, in Regulation of Photosynthesis (eds. Aro E. M., Andersson, B.), Dordrecht: Kluwer, 2001, 395–418.
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Wei, H., Guo, J., Zhang, S. et al. The presence of phosphorylation form of D1 protein in its cross-linked aggregates in high light treated spinach leaves in vivo . CHINESE SCI BULL 51, 69–74 (2006). https://doi.org/10.1007/s11434-005-1529-3
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DOI: https://doi.org/10.1007/s11434-005-1529-3