Populating the equilibrium molten globule state of apomyoglobin under conditions suitable for structural characterization by NMR
Populating the equilibrium molten globule state of apomyoglobin under conditions suitable for structural characterization by NMR
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DOI：
10.1016/s0014-5793(97)01256-8
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发表时间：
1997-11-03
期刊：
影响因子：
3.5
通讯作者：
Wright, PE
中科院分区：
文献类型：
--
作者：
Eliezer, D;Jennings, PA;Wright, PE
确定了去肌红蛋白的平衡熔融球状状态稳定并保持单体状态的条件，这一时间段足以应用三维异核核磁共振实验。初始二维核磁共振谱的质量表明，在这些条件下，可以对大多数蛋白质的共振进行序列特定的指定。用二维H-1-N-15相关实验对蛋白质进行pH滴定，表明平衡中间体在化学位移时间尺度上与展开状态发生快速交换，与天然状态发生中间时间尺度交换，并提出了一种辅助主链共振归属的策略。所描述的条件和技术可应用于其他平衡折叠中间体的表征。(C)1997年欧洲生化学会联合会。
Conditions have been determined under which the equilibrium molten globule state of apomyoglobin is stable and remains monomeric for periods of time sufficient for the application of three-dimensional heteronuclear NMR experiments. The quality of initial two-dimensional NMR spectra suggests that sequence-specific assignments can be made for a majority of the protein resonances under these conditions. A pH titration of the protein followed using two-dimensional H-1-N-15 correlation experiments indicates that the equilibrium intermediate undergoes fast exchange on the chemical shift time scale with the unfolded state and intermediate time scale exchange with the native state, and suggests a strategy to assist with backbone resonance assignments. The conditions and techniques described may be applicable to the characterization of other equilibrium folding intermediates.(C) 1997 Federation of European Biochemical Societies.