MOLECULAR MECHANISM OF REDOX REGULATION OF CHLOROPLAST ATP SYNTHSAE

叶绿体ATP合成酶氧化还原调控的分子机制

• 批准号: 13440238
• 负责人: HISABORI Toru
• 金额: $ 8.26万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13440238/
• 关键词: SWITCH REGION; γSUBUNIT; ATP SYNTHASE; CHLOROPLAST; REDOX REGULATION; εSUBUNIT

In this project, we intended to understand the redox regulation of chloroplast ATP synthase at the molecular level. For these three research years, we obtained the several important findings as follows:A.The switch region from the γ subunit of the redox-sensitive chloroplast F1 was introduced into the bacterial F1 and the redox switching of the rotation was revealed. We revealed that the suppressed enzymatic activity of the oxidized Fl was characterized by more frequent long pauses in the rotation behaviour.B.The regulatory function of the γ and ε subunits of chloroplast ATP synthase was studied using the membrane integrated chimeric complex. We concluded that the interaction between these subunits is important for the redox regulation of FoF1 complex by the γ subunit, and a certain structural matching between these regulatory subunits and the catalytic core of the enzyme must be required to confer the redox regulation mechanism to the bacterial FoF1.C.The sensitivity against tentoxin, the specific inhibitor peptide for the chioroplast F1, was conferred to the bacteria Fl by a single amino acid substitution based on the three dimensional structure analysis. Then the rotation of this mutant Fl was investigated in the presence of this inhibitor. The change of the pauses induced by the inhibitor was revealed.

本项目旨在从分子水平上了解叶绿体ATP合酶的氧化还原调控。在这三年的研究中，我们获得了以下几个重要的发现：A.将氧化还原敏感叶绿体F1的γ亚基的开关区导入细菌F1，揭示了旋转的氧化还原开关。我们揭示了氧化的F1的酶活性被抑制的特征在于旋转行为中更频繁的长暂停。B.利用膜整合嵌合复合物研究了叶绿体ATP合酶γ和ε亚基的调节功能。这些亚基之间的相互作用对于γ亚基对FoF1复合物的氧化还原调节是重要的，而这些调节亚基与酶的催化核心之间必须有一定的结构匹配才能赋予细菌FoF1.C氧化还原调节机制。通过基于三维结构分析的单个氨基酸取代赋予细菌F1。然后在该抑制剂存在下研究该突变体F1的旋转。揭示了抑制剂引起的停顿的变化。

项目成果

Paviova, P., Shimabukuro, K., Hisabori, T., Groth, G., Lill, H., Bald, D.: "Complete inhibition and partial re-activation of single F1-ATPase molecules by tentoxin : New properties of the re-activated enzyme"J.Biol.Chem.. 279(11). 9685-9688 (2004)

Paviova, P.、Shimabukuro, K.、Hisabori, T.、Groth, G.、Lill, H.、Bald, D.：“腱毒素对单个 F1-ATPase 分子的完全抑制和部分重新激活：

Sugiyama, K., Hisabori, T.: "Conformational change of the chloroplast ATP synthase on the enzyme activation process detected by the trypsin sensitivity of the gamma subunit"Biochem.Biophys.Res.Commun.. 301(2). 311-316 (2003)

Sugiyama, K.、Hisabori, T.：“通过 γ 亚基的胰蛋白酶敏感性检测到的叶绿体 ATP 合酶对酶激活过程的构象变化”Biochem.Biophys.Res.Commun. 301(2)。

Bald D, Noji H, Yoshida M, Hirono-Hara Y, Hisabori T.: "Redox regulation of the rotation of F_1-ATP synthase."J Biol Chem.. 276. 39505-39507 (2001)

Bald D、Noji H、Yoshida M、Hirono-Hara Y、Hisabori T.：“F_1-ATP 合酶旋转的氧化还原调节。”J Biol Chem.. 276. 39505-39507 (2001)

Groth G, Hisabori T, Lill H, Bald D.: "Substitution of a single amino acid switches the tentoxin-resistant thermophilic F1-ATPase into a tentoxin-sensitive enzyme"J.Biol.Chem.. 277(23). 20117-20119 (2002)

Groth G、Hisabori T、Lill H、Bald D.：“单个氨基酸的取代可将抗腱毒素的嗜热 F1-ATP 酶转变为对腱毒素敏感的酶”J.Biol.Chem.. 277(23)。

Sugiyama, K., Hisabori, T.: "Conformational change of the chloroplast ATP synthase on the enzyme activation process detected by the trypsin sensitivity of the γ subunit"Biochem.Biophys.Res.Commun.. 301. 311-316 (2003)

Sugiyama, K., Hisabori, T.：“通过 γ 亚基的胰蛋白酶敏感性检测到的酶激活过程中叶绿体 ATP 合酶的构象变化”Biochem.Biophys.Res.Commun.. 301. 311-316 (2003)