Relation between the structure and function of proteins studied by the changes of tyrosine and tryptophan residues.

通过酪氨酸、色氨酸残基的变化研究蛋白质结构与功能的关系。

• 批准号: 14570103
• 负责人: NAGAI Masako
• 金额: $ 2.18万
• 依托单位: Hosei University (2004)Kanazawa University (2002-2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570103/
• 关键词: hemoglobin; SH3; near-UV CD; UV resonance Raman; mutants; tyrosine; tryptophan; quaternary structure transition; 高次構造変化; 芳香族アミノ酸; 酸素結合機能; CD; 共鳴ラマン; HbM; アロステリックエフェクター; 蛋白質の機能; 円二色性(CD); PI3K; 異常血色素; SrcSH3 domein; 共鳴ラマン分光; 円二色性

To get insight into how the quaternary structure changes correlate to their functions, we examined the near-UV CD and UV resonance Raman spectra of hemoglobin and the domein of Src-homology-3 protein with and without ligands. Using four newly synthesized mutant hemoglobins at α42Tyr,α140Tyr,β145Tyr, and/or β37Trp, it was clarified that the main contributors for a negative CD band, a T-sate marker band, are α140Tyr and β145Tyr, located at C-terminal positions. The T-structure specific UV resonance Raman bands of Tyr and Trp were characterized using a natural mutant Hb, Hb M Boston and a Ni-Fe Hybrid hemoglobin.Src-homology-3(SH3) protein recognize a Pro rich peptides and communicate with the other proteins. We demonstrated that SH3 interacts to the ligand peptide via Tyr residue(s) using UV CD and UV resonance Raman spectoscopy. SH3 has six Tyr residues. Specific Tyr residue for the interaction with Pro-rich peptide was specified as 14Tyr using mutants synthesized in E.coli each Tyr replaced by Ala. Interestingly, Src-SH3 and PI3K-SH3 showed different shtructure changes with the interaction of ligand peptides reflecting the different protein recognition.

为了深入了解四级结构的变化如何与它们的功能相关联，我们研究了血红蛋白和Src-同源性-3蛋白的domein在有和没有配体的情况下的近紫外CD和UV共振拉曼光谱。使用新合成的α 42 Tyr、α 140 Tyr、β 145 Tyr和/或β 37 Trp突变血红蛋白，阐明了阴性CD带（T状态标记带）的主要贡献者是位于C末端位置的α 140 Tyr和β 145 Tyr。利用天然突变血红蛋白Hb M Boston和Ni-Fe Hybrid血红蛋白研究了Tyr和Trp的T-结构特异性紫外拉曼共振谱带。利用紫外CD和紫外共振拉曼光谱证明了SH 3通过Tyr残基与配体肽相互作用。SH 3具有六个Tyr残基。使用在大肠杆菌中合成的突变体，将与富含Pro的肽相互作用的特定Tyr残基指定为14 Tyr，每个Tyr被Ala取代。有趣的是，Src-SH 3和PI 3 K-SH 3在与配体肽相互作用时显示出不同的结构变化，反映了不同的蛋白质识别。

项目成果

Jin, Y., Sakurai, H., Nagai, Y., Nagai, M.: "Changes of near-UV CD spectrum of human hemoglobin upon oxygen binding"Biospectroscopy. (In press).

Jin, Y.、Sakurai, H.、Nagai, Y.、Nagai, M.：“氧结合时人血红蛋白近紫外 CD 光谱的变化”生物光谱。

Heme structures of five variants of hemoglobin M probed by resonance Raman spectroscopy.

• DOI: 10.1021/bi036170g
• 发表时间: 2004-06
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Yayoi Jin;M. Nagai;Y. Nagai;S. Nagatomo;T. Kitagawa

Changes in the abnormal α-subunit upon CO-binding to the normal β-subunit of Hb M Boston : resonance Raman, EPR, and CD study.

与 Hb M Boston 的正常 β 亚基共同结合后异常 α 亚基的变化：共振拉曼、EPR 和 CD 研究。

• 发表时间: 2002
• 期刊: Biophysical Chemistry 98
• 作者: Nagatomo;S.;Jin;Y.;Nagai;M.;Hori;H.;Kitagawa;T.
• 通讯作者: T.

Differences in changes of the α1β2 subunit contacts between ligand binding to the α and β subunits of Hb A. UV resonance Raman analysis using Ni-Fe hybrid hemoglobin

与 Hb A 的 α 和 β 亚基结合的配体之间 α1β2 亚基接触变化的差异。使用 Ni-Fe 杂化血红蛋白进行 UV 共振拉曼分析

• 发表时间: 2002
• 期刊: Biochemistry 41
• 作者: Nagatomo;S.;Nagai;M.;Shibayama;N.;Kitagawa;T.
• 通讯作者: T.

Quaternary structures of intermediately liganded human hemoglobin A and influences from strong allosteric effectors ; resonance Raman investigation.

中间配体人血红蛋白A的四级结构和强变构效应子的影响；

• 期刊: (投稿中)
• 作者: Nagatomo;S.;Nagai;M.;Mizutani;Y.;Yonetani;T.;Kitagawa;T.
• 通讯作者: T.