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学科主题: 天然产物有机化学
题名: 细胞色素b5突变体V45H的初步研究
其他题名: NMR preliminary study on cytochrome b5 mutant V45H
作者: 曹春阳 ; 张乃霞 ; 王跃丰 ; 吴厚铭 ; 王志强 ; 王韵华 ; 黄仲贤
通讯作者: 吴厚铭
刊名: 化学学报
发表日期: 2001-01-01
卷: 59, 期:4, 页:578-586
收录类别: SCI
部门归属: 中国科学院上海有机化学研究所生命有机化学实验室;复旦大学化学系
摘要: 通过分析在H^20和D^2O中采集的 DQF—COSY,TOCSY和NOESY等二维核磁共振波谱鉴定了细胞色素b5定点突变体V45H(残基 val~45突变为Hi~45)的大多数氨基酸残基的质子自旋系统.通过解析 NOESY谱中的DNN(I,I,+1),d^aN(I,I+1), daN(I,I+2), dan(I,I+3), daβ(I,I+3)和dβN(I,I+1)等NOE相关,完成了其序列特异性归属以及主链和侧链质子共振信号的全归属.突变体V45H的二级结构分析表明残基Val~45突变为His~45对分子的整体折叠影响不大.但是,与野生型细胞色素b5相比较,突变体 V45H主链酰胺质子的化学位移指数提示突变使其血红素疏水腔的微环境受到扰动.以上实验结果为进一步测定V45H的溶液结构和分析残基 vla~45在蛋白质中的作用提供了基础
英文摘要: In this paper, most of spin systes forspin sstems for amino acid residues in V45H, a site-directed mutant of cytochrome b5 where the residue Val~45, have been idntified trough analysis of 2D NMR ~1H-~1H DQF-COSY,TOCSY and NOESY spectra acquired in H^2O and D^O. The esquence-specific assignment of spin systems is obtained by Dnn(I,I+1), daN(I,I-1), daN(I,I+2), daN(I,I+3), daβ(I,I+3) and dβN(I,I+1) NOEs correlation found in NOESY spectra and the complete assignment of proton resonances for the backbone and side chain have been achieved. The analysis of secondary structure of V45H revelas that the mutation from Val~45 has little effect in the global folding of the protein. However, the chemical shift index of amide protons of V45H in comparison to those of wild type cytochrome b5 indicates that the hemem pocket environment is disturbed by the mutation. These experimental results provide a basis for further determination of the solution structure of V45H and the roles of residue Val!45 in the protein.
语种: 中文
内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/17291
Appears in Collections:上海有机化学研究所_期刊论文

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Recommended Citation:
曹春阳,张乃霞,王跃丰,等. 细胞色素b5突变体V45H的初步研究[J]. 化学学报,2001,59(4):578-586.
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