Molecular Approach to the Regulatory Mechanism of Chloroplast ATP Synthase

叶绿体 ATP 合酶调控机制的分子方法

• 批准号: 08640822
• 负责人: HISABORI Toru
• 金额: $ 1.41万
• 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 (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08640822/
• 关键词: ATP Synthase; gamma subunit; thioredoxin; Conformational Change; epsilon subunit; チナレドキシン

We have studied the change of the catalytic activity of chimeric complexes which were formed by CF_1-gamma, and alpha and beta subunits of thermophilic bacterial F_1 after formation or reduction of the disulfide bridge of different gamma subunits modified by oligonucleotide-directed mutagenesis techniques. For this purpose, three mutant gamma subunits were produced : gamma_<DELTA194-230>, here 37 amino acids from Pro-194 to Ile-230 are deleted, gamma_<C199A>, Cys-199 is changed to Ala, and gamma_<DELTA200-204>, amino acids from Asp-200 to Lys-204 are deleted. All of the chimeric subunit complexes produced from each of these mutant CF_1-gamma subunits and alpha and beta subunits from thermophilic bacterial F_1 lost the sensitivity against thiol reagents when compared to the complex containing wild type CF_1-gamma. The pH optimum (pH 8.5 to pH 9.0) and the concentration of methanol to stimulate ATPase activities were not affected by these mutations. These indicate that the introduction o … More f the mutations did not change the main features of ATPase activity of the chimeric complex.However, the interaction between gamma subunit and epsilon subunit was strongly influenced by the type of gamma subunit itself. Although the ATPase activity of the chimeric complex which contained gamma_<DELTA200-204> or gamma_<C199A> was inhibited by the addition of recombinant epsilon subunit from CF_1 similarly to complexes containing the reduced wild-type gamma subunit, the recombinant epsilon subunit did not inhibit the ATPase of the complex which contained the oxidized form of gamma subunit, suggesting the affinity of the epsilon subunit to the gamma subunit is dependent on the state of the gamma subunit or the epsilon subunit may bind to the oxidized form of gamma subunit in a mode that does not inhibit the activity. The ATPase activity of the complex which contains gamma_<DELTA194-230> was not efficiently inhibited by epsilon subunit. These results show that the formation or the reduction of the disulfide bond on the gamma subunit may induce a conformational change in the region which directly affects the interaction of this subunit with the adjacent epsilon subunit. Less

我们研究了嗜热菌F_1的CF_1-γ、α和β亚基经三肽定向突变技术修饰后，形成或还原不同γ亚基的二硫键后，形成的嵌合复合物的催化活性的变化。为此目的，产生了三种突变体γ亚基：γ_<DELTA194-230>，这里从Pro-194到Ile-230的37个氨基酸缺失，γ_<C199A>，Cys-199变为Ala，并且γ_<DELTA200-204>，从Asp-200到Lys-204的氨基酸缺失。与含有野生型CF_1-γ的复合物相比，由这些突变CF_1-γ亚基和来自嗜热细菌F_1的α和β亚基中的每一个产生的所有嵌合亚基复合物都丧失了对巯基试剂的敏感性。最适pH值（pH 8.5至pH 9.0）和甲醇的浓度，以刺激ATP酶的活性不受这些突变。这些表明， ...更多信息 突变并没有改变嵌合复合物ATP酶活性的主要特征，但γ亚基与ATP酶亚基之间的相互作用受到γ亚基类型的强烈影响。虽然含有γ_或γ_的嵌合复合物的ATP酶活性<DELTA200-204><C199A>被来自CF_1的重组α-亚基的加入所抑制，类似于含有还原的野生型γ亚基的复合物，但重组α-亚基不抑制含有氧化形式的γ亚基的复合物的ATP酶，这表明γ亚基对γ亚基的亲和力取决于γ亚基的状态，或者γ亚基可以结合γ亚基的氧化形式，γ亚基以不抑制活性的方式存在。含γ_3的复合物的ATP酶活性<DELTA194-230>不受γ_3亚基的有效抑制。这些结果表明，γ亚基上二硫键的形成或还原可能诱导该区域的构象变化，这直接影响该亚基与相邻的γ亚基的相互作用。少

项目成果

TORU HISABORI 他5名: "The regulatory functions of the γ and ε subunits from chloroplast CF_1 are transferred to the core complex,α_3β_3,from thermophilic bacterial F_1" Eur.J.Biochem.247. 1158-1165 (1997)

TORU HISABORI 和其他 5 人：“叶绿体 CF_1 的 γ 和 ε 亚基的调节功能从嗜热细菌 F_1 转移到核心复合物 α_3β_3”Eur.J.Biochem.247 (1997)。

Hisabori, T., Kato, Y., Motohashi, K., Kroth-Pancic, P., Strotmann, H., and Amano, T.: "The Regulatory Functions of the gamma and epsilon Subunits from Chloroplast CF_1 are Transferred to the Core Complex, alpha_3beta_3, from Thermophilic Bacterial F_1" E

Hisabori, T.、Kato, Y.、Motohashi, K.、Kroth-Pancic, P.、Strotmann, H. 和 Amano, T.：“来自叶绿体 CF_1 的 γ 和 ε 亚基的调节功能被转移到

ToRu HISABORI,他5名: "The regulatory functions of the γand ε subunits from chloroplast CF_1 are transferred to the core complex,α_3β_3,from thermophilic bacterial F_1" Eur.J.Biochem.247. 1158-1165 (1997)

ToRu HISABORI 和其他 5 人：“叶绿体 CF_1 的 γ 和 ε 亚基的调节功能从嗜热细菌 F_1 转移到核心复合物 α_3β_3”Eur.J.Biochem.247 (1997)。