Towards automated screening of two-dimensional crystals

被引:28
作者
Cheng, Anchi
Leung, Albert
Fellmann, Denis
Quispe, Joel
Suloway, Christian
Pulokas, James
Abeyrathne, Priyanka D.
Lam, Joseph S.
Carragher, Bridget
Potter, Clinton S.
机构
[1] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA
[2] Univ Guelph, Dept Mol & Cellular Biol, Guelph, ON N1G 2W1, Canada
关键词
TEM; 2D crystallization; automation; membrane proteins; electron crystallography;
D O I
10.1016/j.jsb.2007.09.012
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
Screening trials to determine the presence of two-dimensional (2D) protein crystals suitable for three-dimensional structure determination using electron crystallography is a very labor-intensive process. Methods compatible with fully automated screening have been developed for the process of crystal production by dialysis and for producing negatively stained grids of the resulting trials. Further automation via robotic handling of the EM grids, and semi-automated transmission electron microscopic imaging and evaluation of the trial grids is also possible. We. and others, have developed working prototypes for several of these tools and tested and evaluated them in a simple screen of 24 crystallization conditions. While further development of these tools is certainly required for a turn-key system, the goal of fully automated screening appears to be within reach. (c) 2007 Elsevier Inc. All rights reserved.
引用
收藏
页码:324 / 331
页数:8
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