Thielmann Yvonne, Koepke Juergen, Michel Hartmut
Department of Molecular Membrane Biology, Max-Planck-Institute of Biophysics, Max-von-Laue Str. 3, 60438, Frankfurt, Germany.
J Struct Funct Genomics. 2012 Jun;13(2):63-9. doi: 10.1007/s10969-011-9118-y. Epub 2011 Nov 19.
Structure determination of membrane proteins and membrane protein complexes is still a very challenging field. To facilitate the work on membrane proteins the Core Centre follows a strategy that comprises four labs of protein analytics and crystal handling, covering mass spectrometry, calorimetry, crystallization and X-ray diffraction. This general workflow is presented and a capacity of 20% of the operating time of all systems is provided to the European structural biology community within the ESFRI Instruct program. A description of the crystallization service offered at the Core Centre is given with detailed information on screening strategy, screens used and changes to adapt high throughput for membrane proteins. Our aim is to constantly develop the Core Centre towards the usage of more efficient methods. This strategy might also include the ability to automate all steps from crystallization trials to crystal screening; here we look ahead how this aim might be realized at the Core Centre.
膜蛋白及膜蛋白复合物的结构测定仍然是一个极具挑战性的领域。为推动膜蛋白研究工作，核心中心采用了一种策略，该策略涵盖四个蛋白质分析与晶体处理实验室，涉及质谱分析、量热法、结晶及X射线衍射。本文介绍了这一通用工作流程，并在欧洲科学基金会研究基础设施战略论坛（ESFRI）的Instruct计划框架内，将所有系统20%的运行时间提供给欧洲结构生物学界。文中给出了核心中心提供的结晶服务的描述，包括筛选策略、所用筛选方法以及为适应膜蛋白高通量研究而做出的调整等详细信息。我们的目标是不断推动核心中心采用更高效的方法。该策略可能还包括实现从结晶试验到晶体筛选所有步骤自动化的能力；在此，我们展望这一目标在核心中心如何实现。
