Structual formation and structure-function relationship of protein studied with the use peptides

用肽研究蛋白质的结构形成和结构-功能关系

• 批准号: 16550149
• 负责人: HIROTA Shun
• 金额: $ 2.37万
• 依托单位: Kyoto Pharmaceutical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16550149/
• 关键词: protein; peptide; structure-function relationship; cytochrome c; plastocyanin; tyrosine-containing peptide; electron transfer; 構造形成; 構造ー機能相関; シトオクロムc

In biosystems, proteins interact with each other. In this research, oligopeptides are used as models of the protein interacting site, and the interaction between the protein and the peptide is studied in detail. The following results are obtained.1. Cytochrome c(cyt c) was reduced by a tyrosine-containing peptide, tyrosyltyrosylphenylalanine (TyrTyrPhe), at pH 6.0-8.0. Cyt c was reduced at high peptide concentration, whereas the reaction did not occur effectively at low concentration. The reciprocal initial rate constant (1/k_<int>) increased linearly against the reciprocal peptide concentration and against the linear proton concentration, whereas log k_<int> decreased linearly against the root of the ionic strength. These results show that deprotonated (TyrTyrPhe)., presumably deprotonated at a tyrosine site, reduces cyt c by formation of an electrostatic complex.2. From the MALDI-TOF MS spectra of the reaction products obtained from the reaction between cyt c and YYF, formation of a … More quinone and other tyrosine derivatives of the peptide was supported. These products should have been produced from a tyrosyl radical. The results are interpreted that a cyt c_<ox>/(TyrTyrPhe)【tautomer】 cyt c_<red>/(TyrTyrPhe)・ equilibrium is formed, which is usually shifted to the left. This equilibrium may shift to the right by reaction of the produced tyrosyl radical with the tyrosine sites of unreacted TvrTvrPhe peptides.3. Oxidized plastocyanin (PC) was reduced with TyrTyrTyr at neutral pH. The reciprocal initial rate constant (1/k_<int>) increased linearly with the reciprocal TyrTyrTyr concentration and proton concentration. The results showed that PC was reduced by the deprotonated species of TyrTyrTyr. A linear increase of log k_<int> with increase in the ionic strength was observed due to decrease in the electrostatic repulsion between negatively charged PC and deprotonated (TyrTyrTyr).These results show that a metalloprotein could be reduced at neutral pH with a tyrosine-containing oligopeptide. Less

在生物系统中，蛋白质相互作用。本研究以寡肽作为蛋白质相互作用位点的模型，详细研究了蛋白质与寡肽之间的相互作用。得到以下结果.细胞色素c（cyt c）在pH 6.0 - 8.0下被含酪氨酸的肽酪氨酰酪氨酰苯丙氨酸（TyrTyrPhe）还原。Cyt c在高肽浓度下被还原，而反应在低浓度下不有效地发生。初始速率常数的倒数（1/k_）随肽浓度的倒数和质子浓度的线性增加而线性增加，而log k_随离子强度的根线性减小。<int><int>这些结果表明去质子化的（TyrTyrPhe），推测在酪氨酸位点去质子化，通过形成静电复合物还原CytC。从从cyt c和YYF之间的反应获得的反应产物的MALDI-TOF MS光谱， ...更多信息 负载肽的醌和其它酪氨酸衍生物。这些产物应该是由酪氨酰基自由基产生的。结果表明，形成了cyt c_/（TyrTyrPhe）[互变异构体] cyt c_/（TyrTyrPhe）·平衡，该平衡通常向左移动。<ox><red>通过产生的酪氨酰基自由基与未反应的TvrTvrPhe肽的酪氨酸位点的反应，该平衡可向右移动。在中性pH条件下，TyrTyrTyr还原氧化质体蓝素（PC）的反应速率常数（1/k）与TyrTyrTyr浓度和质子浓度的倒数呈线性关系。<int>结果表明，PC被TyrTyrTyr的去质子化物质还原。由于带负电荷的PC与去质子化的（TyrTyrTyr）之间的静电排斥力减小，log k_（？）随离子强度的增加而线性增加，表明含酪氨酸的寡肽在中性pH下可以还原金属蛋白。<int>少

项目成果

Interaction of Plastocyanin with Oligopeptides : Effect of Lysine Distribution within the Peptide

质体蓝素与寡肽的相互作用：肽内赖氨酸分布的影响

• 发表时间: 2004
• 期刊: J.Inorg.Biochem. 98
• 作者: S.Hirota;H.Okumura;S.Arie;K.Tanaka;M.Shionoya;T.Takabe;N.Funasaki;Y.Watanabe
• 通讯作者: Y.Watanabe

Activation process of [NiFe] hydrogenase elucidated by high-resolution X-ray analyses: Conversion of the ready to the unready state

• DOI: 10.1016/j.str.2005.07.018
• 发表时间: 2005-11-01
• 期刊: STRUCTURE
• 影响因子: 5.7
• 作者: Ogata, H;Hirota, S;Higuchi, Y
• 通讯作者: Higuchi, Y

Reduction of ferricytochrome c by tyrosyltyrosylphenylalanine

酪氨酰酪氨酰苯丙氨酸还原铁细胞色素 c

• 发表时间: 2005
• 期刊: J. Biol. Inorg. Chem. 10・4
• 作者: S.Hirota;H.Okumura;T.Kondoh;N.Funasaki;T.Takabe;Y.Watanabe;S.Hirota et al.
• 通讯作者: S.Hirota et al.

Reduction of plastocyanin by tyrosine-containing oligopeptides.

含酪氨酸寡肽减少质体蓝素。

• 发表时间: 2006
• 期刊: J. Inorg. Biochem 100
• 作者: Hirota S;Okumura H;Kondoh T;Funasaki N;Takabe T;Watanabe Y
• 通讯作者: Watanabe Y

Conformational changes during apoplastocyanin fokding observed by photocleavable modification and transient grating

通过光裂解修饰和瞬态光栅观察质外体蓝蛋白形成过程中的构象变化

• 发表时间: 2006
• 期刊: J.Am.Chem.Soc. 128・23
• 作者: S.Hirota;Y.Fujimoto;J.Choi;N.Baden;N.Katagiri;M.Akiyama;R.Hulsker;M.Ubbink;T.Okajima;T.Takabe;N.Funasaki;Y.Watanabe;M.Terazima
• 通讯作者: M.Terazima