一个17.6kDa蛋白质的高分辨率固态NMR结构
Ivano Bertini1, Anusarka Bhaumik, Gaël De Paëpe
1Magnetic Resonance Center, CERM, University of Florence, Via L. Sacconi, 6-50019 Sesto Fiorentino, Italy. bertini@cerm.unifi.it
Journal of the American Chemical Society
|January 1, 2010
概括
这项研究引入了伪接触转移,从对磁性金属离子到固态NMR来分配蛋白质信号和计算结构. 这种方法为金属蛋白酶12域产生了777个距离限制,实现了高分辨率的结构确定.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 生物物理学的生物物理.
背景情况:
- 固态NMR对于确定蛋白质结构至关重要,但信号分配可能具有挑战性.
- 磁性金属离子可以通过伪接触转移提供远程结构信息.
研究的目的:
- 开发和验证一种使用伪接触转移的策略,用于在固态NMR中明确的信号赋值.
- 为了使精确的距离限制提取用于蛋白质结构计算.
主要方法:
- 在微晶蛋白样本中使用了对磁性金属离子 () 替代.
- 采用了各种固态NMR实验,包括PDSD,DARR,CHHC,PAR和PAIN-CP.
- 获得伪接触转移来导出距离限制.
主要成果:
- 为矩阵金属蛋白酶12的催化域生成了777个明确的距离限制 (序列,中程,远程).
- 确定了一个高分辨率的蛋白质结构,骨干RMSD为1.0 ± 0.2 Å.
- 计算的结构与现有的X射线结构 (RMSD为1.3 Å) 有很好的一致性.
结论:
- 从磁性离子的伪接触转移对于固态NMR中明确的信号赋值是有效的.
- 这一策略显著推进了使用固态NMR的蛋白质结构确定.
- 这种方法可能可以将其推广到使用偏磁标签的非金属蛋白.
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