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学科主题: 公共卫生
题名:
Probing the conformational state of a truncated staphylococcal nuclease R using time of flight mass spectrometry with limited proteolysis
作者: Yang, F; Cheng, Y; Peng, JR; Zhou, JM; Jing, GZ
关键词: conformation ; ligand binding ; mass spectrometry ; truncated staphylococcal nuclease
刊名: EUROPEAN JOURNAL OF BIOCHEMISTRY
发表日期: 2001-08-01
卷: 268, 期:15, 页:4227-4232
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
关键词[WOS]: N-TERMINAL FRAGMENTS ; RESIDUAL STRUCTURE ; CRYSTAL-STRUCTURE ; MOLTEN GLOBULE ; PROTEINS ; PEPTIDE ; FOLD
英文摘要:

The conformational state of C-terminally truncated staphylococcal nuclease R (SNR135), with and without bound ligands, has been studied by performing limited proteolysis with a specific endoproteinase Glu-C followed by electrophoresis and mass spectrometry. Comparison of the accessibility of the cleavage sites shows that the C-terminal truncation of 14 amino-acid residues causes significant unfolding of the C-terminal part of alpha helix 1 and the center of alpha helix 2, but there is little effect on other regions of the nuclease, in particular the N-terminal subdomain, which includes the active site of the nuclease. The truncation also makes the overall conformation of the nuclease more loose and flexible. Binding of ligands makes helices 1 and 2 more resistant to protease Glu-C attack and converts the partially unfolded state to a native-like state, although the conformational stability of the SNR135 complex is still much lower than that of the full-length enzyme. The results suggest that the amino-acid residues around the active site in the truncated nuclease are arranged in a similar topology to those in the full-length nuclease. The study shows that there is a clear-cut correlation between protease susceptibility and conformational stability of the protein, and the initial proteolytic events are the most critical for evaluating the conformational features of the protein. This study demonstrates how mass spectrometry can be combined with limited proteolysis to observe conformational changes induced by ligand binding.

语种: 英语
WOS记录号: WOS:000170399300014
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/123622
Appears in Collections:北京大学医药卫生分析中心_期刊论文

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作者单位: 1.Acad Sinica, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China
2.Beijing Univ, Med & Hlth Anal Ctr, Beijing 100871, Peoples R China

Recommended Citation:
Yang, F,Cheng, Y,Peng, JR,et al. Probing the conformational state of a truncated staphylococcal nuclease R using time of flight mass spectrometry with limited proteolysis[J]. EUROPEAN JOURNAL OF BIOCHEMISTRY,2001,268(15):4227-4232.
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