Lysine methylation as a routine rescue strategy for protein crystallization

被引:198
作者
Walter, Thomas S.
Meier, Christoph
Assenberg, Rene
Au, Kin-Fai
Ren, Jingshan
Verma, Anil
Nettleship, Joanne E.
Owens, Raymond J.
Stuart, David I.
Grimes, Jonathan M.
机构
[1] Univ Oxford, Oxford Prot Prod Facil, Oxford OX3 7BN, England
[2] Univ Oxford, Div Struct Biol, Oxford OX3 7BN, England
基金
英国医学研究理事会; 英国惠康基金;
关键词
D O I
10.1016/j.str.2006.09.005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Crystallization remains a critical step in X-ray structure determination. Because it is not generally possible to rationally predict crystallization conditions, commercial screens have been developed which sample a wide range of crystallization space. While this approach has proved successful in many cases, a significant number of proteins fail to crystallize despite being soluble and monodispersed. It is established that chemical modification can facilitate the crystallization of otherwise intractable proteins. Here we describe a method for the reductive methylation of lysine residues which is simple, inexpensive, and efficient, and report on its application to ten proteins. We describe the effect of methylation on the physicochemical properties of these proteins, and show that it led to diffraction-quality crystals from four proteins and structures for three that had hitherto proved refractory to crystallization. The method is suited to both low- and high-throughput laboratories.
引用
收藏
页码:1617 / 1622
页数:6
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