Zacharchenko Thomas, Wright Stephanie
School of Biology and Astbury Centre for Structural Molecular Biology, University of Leeds, Woodhouse, Leeds LS2 9JT, United Kingdom.
IUCrJ. 2021 Jan 11;8(Pt 2):154-160. doi: 10.1107/S2052252520015754. eCollection 2021 Mar 1.
The production of diffraction-quality protein crystals is challenging and often requires bespoke, time-consuming and expensive strategies. A system has been developed in which the BCL6 BTB domain acts as a crystallization chaperone and promiscuous assembly block that may form the basis for affinity-capture crystallography. The protein of interest is expressed with a C-terminal tag that interacts with the BTB domain, and co-crystallization leads to its incorporation within a BTB-domain lattice. This strategy was used to solve the structure of the SH3 domain of human nebulin, a structure previously solved by NMR, at 1.56 Å resolution. This approach is simple and effective, requiring only routine protein complexation and crystallization screening, and should be applicable to a range of proteins.
生成衍射质量的蛋白质晶体具有挑战性，通常需要定制的、耗时且昂贵的策略。已开发出一种系统，其中BCL6 BTB结构域充当结晶伴侣和混杂组装模块，这可能构成亲和捕获晶体学的基础。感兴趣的蛋白质通过与BTB结构域相互作用的C端标签进行表达，共结晶导致其掺入BTB结构域晶格中。该策略用于解析人伴肌动蛋白SH3结构域的结构，该结构先前通过核磁共振解析，分辨率为1.56 Å。这种方法简单有效，仅需常规的蛋白质复合和结晶筛选，并且应适用于一系列蛋白质。
