Physiological and biochemical studies on leaf development and senescence : Degradation of Rubisco mediated by reactive oxygen species

叶片发育和衰老的生理生化研究：活性氧介导的 Rubisco 降解

• 批准号: 09460036
• 负责人: MAE Tadahiko
• 金额: $ 4.16万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460036/
• 关键词: Rubisco; reactive oxygen species; light stress; oxidative stress; degradation; photosynthesis; 葉緑体; 植物; 酸素ストレス

Rubilose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39, Rubiscso) is a bifunctional enzyme which catalyzes two competing reactions, photosynthetic COィイD22ィエD2 assimilation and photorespiratory carbon oxidation in the stroma of the chloroplasts, and is the most abundant protein in leaves. During leaf senescence or under environmental stress conditions, Rubisco is one of the early proteins that are broken down in leaves. However, only limited information is available on the triggering mechanisms that cause Rubisco degradation in plants. Here, we found that the large subunit of Rubisco is directly, fragmented into the 37-kDa and 16-kDa fragments by reactive oxygen species generated in the illuminated lysates of chloroplasts or intact chloroplasts. Analysis of the terminal amino acid residues of both fragments indicated that the fragmentation of the LSU occurs at Gly-329. Purified Rubisco, exposed to a hydroxyl radical generating system, was also cleaved at the same site. Reactive oxygen species generated at its catalytic site by a Fenton-type reaction may trigger the site specific degradation of the LSU in the light.

Rubilose-1，5-bisphosphate carboxylase/oxygenase（EC 4.1.1.39，Rubiscso）是一种双功能酶，其催化两种竞争反应，光合作用CO2 D22同化和叶绿体基质中的光呼吸碳氧化，并且是叶中最丰富的蛋白质。在叶片衰老或环境胁迫条件下，Rubisco是叶片中被分解的早期蛋白质之一。然而，只有有限的信息是在植物中引起Rubisco降解的触发机制。在这里，我们发现，大亚基的Rubisco是直接的，分裂成37-kDa和16-kDa的片段由活性氧物种中产生的叶绿体或完整的叶绿体的裂解物的照明。两个片段的末端氨基酸残基的分析表明，LSU的片段发生在Gly-329。纯化的Rubisco，暴露于羟基自由基生成系统，也被切割在相同的网站。在其催化位点通过芬顿型反应产生的活性氧物质可引发光中LSU的位点特异性降解。

项目成果

Makino, A., and Mae, T.: "Nitrogen partitioning to ribulose-1,5-bisphosphate and chlorophyll in the leaves of rice grown under different irradiances, temperatures, and partial pressures of COィイD22ィエD2. In "Proceedings of the 5ィイD1thィエD1 International Symp

Makino, A. 和 Mae, T.：“在不同辐照度、温度和 CO2D22 分压下生长的水稻叶片中，氮分配为 1,5-二磷酸核酮糖和叶绿素。”第 5 届 D1thD1 国际 Symp 会议记录

Ishida, H., Makino, A., and Mae, T.: "Detection of the cleavage site of the large subunit of Rubisco by active oxygen species and its inhibition by CABP in the lysates of wheat chloroplasts. In "Proceedings of the XIth International Congress on Photosynth

Ishida, H.、Makino, A. 和 Mae, T.：“通过活性氧物质检测 Rubisco 大亚基的裂解位点，并在小麦叶绿体裂解物中通过 CABP 对其进行抑制。”《第 XI 届会议论文集》

Ishida H.: "Fragmentation of the large subumit of ribulose-1,5-bisphosphate carboxylase by active oxygen species occurs near Gly-329"J.Biol.Chem.. 274・8. 5223-5226 (1999)

Ishida H.：“活性氧对核酮糖-1,5-二磷酸羧化酶的大亚基的断裂发生在 Gly-329 附近”J.Biol.Chem.. 274・8 (1999)。

前忠彦: "Rubiscoは光合成に対して過剰存在するか?"化学と生物. 37・2. 113-114 (1999)

前忠彦：“Rubisco 是否过量存在？”化学与生物学 113-114（1999）。

Mae T.: "Leaf senescence and nitrogen metabolism. In"Programmed Cell Death and Related Processes""Ed.by Nooden, L., Academic, Press New York(印刷中). (2000)

Mae T.：“叶衰老和氮代谢。《程序性细胞死亡和相关过程》”，Nooden，L. 编，纽约学术出版社（2000 年出版）。