Membrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK; Research Complex at Harwell, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot OX11 0FA, UK.
Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE, UK.
Methods. 2018 Sep 1;147:150-162. doi: 10.1016/j.ymeth.2018.05.014. Epub 2018 May 18.
Integral membrane proteins are among the most fascinating and important biomolecules as they play a vital role in many biological functions. Knowledge of their atomic structures is fundamental to the understanding of their biochemical function and key in many drug discovery programs. However, over the years, structure determination of integral membrane proteins has proven to be far from trivial, hence they are underrepresented in the protein data bank. Low expression levels, insolubility and instability are just a few of the many hurdles one faces when studying these proteins. X-ray crystallography has been the most used method to determine atomic structures of membrane proteins. However, the production of high quality membrane protein crystals is always very challenging, often seen more as art than a rational experiment. Here we review valuable approaches, methods and techniques to successful membrane protein crystallisation.
整合膜蛋白是最迷人和最重要的生物分子之一，因为它们在许多生物功能中起着至关重要的作用。了解它们的原子结构是理解其生化功能的基础，也是许多药物发现计划的关键。然而，多年来，整合膜蛋白的结构测定被证明远非微不足道，因此它们在蛋白质数据库中的代表性不足。表达水平低、不溶性和不稳定性只是研究这些蛋白质时面临的众多障碍中的一小部分。X 射线晶体学一直是确定膜蛋白原子结构的最常用方法。然而，高质量的膜蛋白晶体的产生总是极具挑战性的，通常更多地被视为一门艺术，而不是一项合理的实验。在这里，我们回顾了成功的膜蛋白结晶的有价值的方法、方法和技术。
