Division of Molecular Biosciences, Membrane Protein Crystallography group, Imperial College London, London SW7 2AZ, UK.
Int J Biochem Cell Biol. 2013 Mar;45(3):636-44. doi: 10.1016/j.biocel.2012.12.018. Epub 2013 Jan 3.
As we appreciate the importance of stabilising membrane proteins, the barriers towards their structure determination are being broken down. This change in mindset comes hand-in-hand with more effort placed on developing methods focused at screening for membrane proteins which are naturally stable in detergent solution or improving those that are not so. In practice, however, it is not easy to decide the best strategy to monitor and improve detergent stability, requiring a decision-making process that can be even more difficult for those new to the field. In this review we outline the importance of membrane protein stability with discussions of the stabilisation strategies applied in context with the use of crystallisation scaffolds and the different types of crystallisation methods themselves. Where possible we also highlight areas that we think could push this field forward with emerging technologies, such as X-ray free electron lasers (X-feL), which could have a big impact on the membrane protein structural biology community. We hope this review will serve as a useful guide for those striving to solve structures of both pro- and eukaryotic membrane proteins.
由于我们认识到稳定膜蛋白的重要性，因此正在克服其结构测定的障碍。这种思维方式的转变伴随着更多的努力，致力于开发针对天然稳定在去污剂溶液中的膜蛋白进行筛选的方法，或改善那些不稳定的膜蛋白的方法。然而，在实践中，决定监测和改善去污剂稳定性的最佳策略并不容易，这需要一个决策过程，对于该领域的新手来说可能更加困难。在这篇综述中，我们概述了膜蛋白稳定性的重要性，并讨论了在结晶支架的应用背景下应用的稳定化策略，以及不同类型的结晶方法本身。在可能的情况下，我们还强调了我们认为可以通过新兴技术推动这一领域发展的领域，例如 X 射线自由电子激光（X-feL），这可能对膜蛋白结构生物学领域产生重大影响。我们希望这篇综述能为那些努力解决原核和真核膜蛋白结构的人提供有用的指导。
