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利用D1蛋白特异性抗体,在强光照(800~2500μmolphotons·m?2·s?1)处理3h的菠菜叶片类囊体膜中检测到4种D1蛋白聚合物,它们的相对含量与处理时光照强度相关.进一步分析表明,70kD聚合物为D1蛋白与CP43交联产物,65和60kD聚合物是D1蛋白与D2蛋白交联产物,41kD聚合物是D1蛋白与细胞色素b559α亚基交联物.1000μmolphotons·m?2·s?1光照处理生成的D1/Cytb559聚合物最多,光强增加时其含量下降.磷酸化苏氨酸抗体Westernblot分析结果显示,D1/Cytb559和D1/CP43聚合物中存在磷酸化蛋白;外加磷酸酯酶处理后,磷酸化的D1/Cytb559和D1/CP43聚合物减少.上述结果表明,强光照处理叶片引起D1蛋白与其他PSⅡ核心蛋白交联,所生成的聚合物中部分D1蛋白以磷酸化形式存在.
Four kinds of D1 protein polymers were detected in spinach leaf thylakoid membranes treated with intense light (800 ~ 2500μmolphotons · m-2 · s · 1) for three hours by using D1 protein-specific antibody. Their relative contents were similar to those of the control Further analysis showed that 70kD polymer was the cross-linked product of D1 protein and CP43, 65 and 60kD polymer was cross-linked product of D1 protein and D2 protein, 41kD polymer was cross-linked with D1 protein and cytochrome b559α subunit. The results showed that D1 / Cytb559 produced the highest concentration of 1000μmolphotons · m · 2 · s · 1, and decreased when the light intensity increased.Western Blot analysis of phospho-threonine antibody showed that D1 / Cytb559 and D1 / CP43 polymers Phosphorylated protein and phosphorylated D1 / Cytb559 and D1 / CP43 polymers decreased with the treatment of phosphatase.The above results indicated that the intense light treatment of the leaves caused the D1 protein to cross-link with other PSII core proteins, Some D1 proteins exist in phosphorylated form.