Rational protein crystallization by mutational surface engineering
Rational protein crystallization by mutational surface engineering
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DOI：
10.1016/j.str.2004.03.008
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发表时间：
2004-04-01
期刊：
影响因子：
5.7
通讯作者：
Derewenda, ZS
中科院分区：
文献类型：
--
作者：
Derewenda, ZS
蛋白质结晶是x射线衍射测定蛋白质结构的一个限制步骤。即使单晶可用，物理质量的不足也可能严重限制可用数据的分辨率，从而限制原子模型的准确性。最近的研究表明，靶向诱变含有大柔性侧链残基的表面斑块，并用较小的氨基酸取代它们，可以有效地制备x射线质量的蛋白质晶体，否则难以结晶。此外，该技术还可用于获得与野生型蛋白质相比质量更好的晶体，有时可将有效分辨率提高1 A或更多。这种新方法最近的几个例子表明，该方法有潜力成为蛋白质晶体学的常规工具。
Protein crystallization constitutes a limiting step in structure determination by X-ray diffraction. Even if single crystals are available, inadequate physical quality may seriously limit the resolution of the available data and consequently the accuracy of the atomic model. Recent studies show that targeted mutagenesis of surface patches containing residues with large flexible side chains and their replacement with smaller amino acids lead to effective preparation of X-ray quality crystals of proteins otherwise recalcitrant to crystallization. Furthermore, this technique can also be used to obtain crystals of superior quality as compared to those grown for the wild-type protein, sometimes increasing the effective resolution by as much as 1 A or more. Several recent examples of this new methodology suggest that the method has the potential to become a routine tool in protein crystallography.