Application of protein engineering to enhance crystallizability and improve crystal properties
Application of protein engineering to enhance crystallizability and improve crystal properties
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DOI：
10.1107/s090744491000644x
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发表时间：
2010-05-01
影响因子：
2.2
通讯作者：
Derewenda, Zygmunt S.
中科院分区：
文献类型：
--
作者：
Derewenda, Zygmunt S.
直到最近，蛋白质结晶大多被认为是一个随机事件，研究人员很少或根本没有控制。随着同步辐射源和探测器的技术进步以及晶体学软件的进步，包括自动化模型构建和验证，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，也有可能对靶蛋白及其复合物进行工程改造，使其更容易形成具有所需X射线衍射质量的晶体。由于目前研究的大多数蛋白质都是通过异源过表达获得的，因此这些技术有望成为常规工具，有可能将经典的结晶筛选转化为更合理的高成功率方法。本文概述了旨在提高结晶性的蛋白质工程方法，并讨论了一些成功应用的例子。
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.