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Early events of folding of β-structure-rich protein

富含β结构的蛋白质折叠的早期事件

基本信息

批准号：
16540373
负责人：
KIHARA Hiroshi
金额：
$ 2.3万
依托单位：
Kansai Medical 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

项目摘要

Protein folds via definite pathway. The purpose of the folding study is to find its folding pathway. We have developed cryo-stopped-flow apparatus with circular dichrosism, x-ray scattering and fluoresecence as probes. We have investigated various types of proteins. Results with alpha-helical protein show the folding rate of alpha-helix formation is so rapid that we cannot measure the rate of alpha-helix formation. In contrast, proteins, mainly consisting of beta-structure, take alpha-helix-rich intrmeditate at its earliest stage. We have focused the folding of SH3, which takes beta-conformation at its native state.SH3 takes alpha-helix-rich intermediate at its earliest stage of folding. We then investigated other beta-structure-rich proteins, and found the intermediate appeared without exception. We compared the amplitude of the burst with other parameters, and found the burst amplitude has good relationship with the helical fraction calculated by a program, Helix2. The program is mainly based on helix-coil transition theory with experimental data of adjacent residues. This suggest that the initial folding cores are formed by the interaction of short-lived alpha-helices.By the one residue mutation of SH3 (Alanine 45 to Glycine), we found the conformation changed from beta-rich to alpha-rich at pH3. This indicates the conversion of beta to alpha occurs more frequently than expected.

蛋白质通过特定途径折叠。折叠研究的目的是寻找其折叠途径。我们研制了以圆二色谱、X射线散射和荧光为探针的低温停流装置。我们研究了各种类型的蛋白质。α-螺旋蛋白的结果显示α-螺旋形成的折叠速率如此之快，以至于我们无法测量α-螺旋形成的速率。相比之下，主要由β结构组成的蛋白质在其最早阶段就具有丰富的α螺旋。我们重点研究了SH 3的折叠过程，SH 3在天然状态下以β-构象存在，在折叠的最早阶段以富含α-螺旋的中间体存在。然后，我们研究了其他富含β结构的蛋白质，发现无一例外地出现了中间体。我们比较了爆发的振幅与其他参数，并发现爆发的振幅与螺旋分数计算程序Helix 2有很好的关系。该程序主要基于螺旋-卷曲转变理论，并结合相邻残基的实验数据。通过SH 3的一个残基突变（丙氨酸45变为甘氨酸），我们发现在pH 3时，构象由富含β构象转变为富含α构象。这表明Beta到Alpha的转换比预期更频繁。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Transient alpha-helical struture during folding of src SH3 domain at subzero temperatures

零下温度下 src SH3 结构域折叠过程中的瞬态 α 螺旋结构

DOI：
发表时间：
期刊：
J. Kansai Medical University in press
影响因子：
0
作者：
Zhijie Qin;Sandip Vyas;Anthony L. Fink;Jinsong Li;Hiroshi Kihara
通讯作者：
Hiroshi Kihara

Transient alpha-helical structure during folding of src SH3 domain at subzero temperatures

零下温度下 src SH3 结构域折叠过程中的瞬态 α 螺旋结构

DOI：
发表时间：
期刊：
J. Kansai Medical University (in press)
影响因子：
0
作者：
Zhijie Qin;Sandip Vyas;Anthony L. Fink;Jinsong Li;Hiroshi Kihara
通讯作者：
Hiroshi Kihara

Structure of Escherichia coli uracil DNA glycosylase and its complexes with nonohydrolyzable substrate analogues in solution observed by synchrotron small-angle X-ray scattering

同步加速器小角X射线散射观察溶液中大肠杆菌尿嘧啶DNA糖基化酶及其与不可水解底物类似物的复合物的结构

DOI：
发表时间：
2006
期刊：
Biofizika 51
影响因子：
0
作者：
Timchenko;S.K.;Kubareva;E.A.;Volkov;E.M.;Voronina;O.L.;Lunin;V.G.;Gonchar;D.A.;Degtiarev;S.K.;Timchenko;M.A.;Kihara;H;Kimura;K
通讯作者：
K

Structure of Escherichia coli uracil DNA glycosylase and its complexes with nonhydrolyzable substrate analogues in solution observed by synchrotron small-angle X-ray scattering

同步加速器小角X射线散射观察溶液中大肠杆菌尿嘧啶DNA糖基化酶及其与不可水解底物类似物的复合物的结构

DOI：
发表时间：
2006
期刊：
Biofizika 51
影响因子：
0
作者：
Timchenko;S.K.;Kubareva;E.A.;Volkov;E.M.;Voronina;O.L.;Lunin;V.G.;Gonchar;D.A.;Degtiarev;S.K.;Timchenko;M.A.;Kihara;H.;Kimura;K.
通讯作者：
K.

Alpha-helical burst in src SH3 folding pathway

src SH3折叠途径中的α螺旋爆发