Derewenda Zygmunt S
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, VA 22908-0736, USA.
Acta Crystallogr D Biol Crystallogr. 2010 May;66(Pt 5):604-15. doi: 10.1107/S090744491000644X. Epub 2010 Apr 21.
Until recently, protein crystallization has mostly been regarded as a stochastic event over which the investigator has little or no control. With the dramatic technological advances in synchrotron-radiation sources and detectors and the equally impressive progress in crystallographic software, including automated model building and validation, crystallization has increasingly become the rate-limiting step in X-ray diffraction studies of macromolecules. However, with the advent of recombinant methods it has also become possible to engineer target proteins and their complexes for higher propensity to form crystals with desirable X-ray diffraction qualities. As most proteins that are under investigation today are obtained by heterologous overexpression, these techniques hold the promise of becoming routine tools with the potential to transform classical crystallization screening into a more rational high-success-rate approach. This article presents an overview of protein-engineering methods designed to enhance crystallizability and discusses a number of examples of their successful application.
直到最近，蛋白质结晶大多被视为一种随机事件，研究者对此几乎没有或完全没有控制权。随着同步辐射源和探测器技术的巨大进步以及晶体学软件（包括自动模型构建和验证）同样令人瞩目的进展，结晶越来越成为大分子X射线衍射研究中的限速步骤。然而，随着重组方法的出现，对目标蛋白质及其复合物进行工程改造，使其更易于形成具有理想X射线衍射质量的晶体也成为可能。由于当今大多数正在研究的蛋白质是通过异源过表达获得的，这些技术有望成为常规工具，有可能将传统的结晶筛选转变为一种更合理、成功率更高的方法。本文概述了旨在提高结晶性的蛋白质工程方法，并讨论了其成功应用的一些实例。
