期刊
JOURNAL OF BIOMOLECULAR NMR
卷 33, 期 2, 页码 95-104出版社
SPRINGER
DOI: 10.1007/s10858-005-2063-y
关键词
inclusion bodies; isotope enrichment; lysozyme; non-native proteins; resonance assignment; tryptophan side chains
A new protocol is described for the isotope (N-15 and C-13,N-15) enrichment of hen egg white lysozyme. Hen egg white lysozyme and an all-Ala-mutant of this protein have been expressed in E. coli. They formed inclusion bodies from which mg quantities of the proteins were purified and prepared for NMR spectroscopic investigations. (HC)-H-1-C-13 and N-15 main chain resonances of disulfide reduced and S-methylated lysozyme were assigned and its residual structure in water pH 2 was characterized by chemical shift perturbation analysis. A new NMR experiment has been developed to assign tryptophan side chain indole resonances by correlation of side chain and backbone NH resonances with the C-gamma supercript stop resonances of these residues. Assignment of tryptophan side chains enables further residue specific investigations on structural and dynamical properties, which are of significant interest for the understanding of non-natives states of lysozyme stabilized by hydrophobic interactions between clusters of tryptophan residues.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据