Derewenda Zygmunt S
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, VA 22908 USA.
Structure. 2004 Apr;12(4):529-35. doi: 10.1016/j.str.2004.03.008.
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.
蛋白质结晶是通过X射线衍射确定结构的一个限制步骤。即使有单晶可用，物理质量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，对含有具有大柔性侧链残基的表面斑块进行定向诱变，并用较小的氨基酸取代它们，可有效制备原本难以结晶的蛋白质的X射线质量晶体。此外，与野生型蛋白质生长的晶体相比，该技术还可用于获得更高质量的晶体，有时有效分辨率可提高多达1埃或更多。这种新方法的几个最新例子表明，该方法有可能成为蛋白质晶体学中的常规工具。
