CRYSTALLOGRAPHIC ANALYSES OF (MOSTLY) METALLOPROTEINS AND MEMBRANE PROTEINS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们希望从介导分子、电子和信息穿过膜双层的膜蛋白晶体中收集x射线衍射数据。我们先前已经确定了结核分枝杆菌的门控大电导机械敏感通道（MscL）的结构，MscL是一种原核通道，其功能是渗透压安全阀（Chang等，Science 282，2220（1998）），和延胡索酸还原酶（FRD）的结构，当延胡索酸是末端电子受体时，FRD催化厌氧呼吸的最后一步（艾弗森等人，Science 284，1961（1999））。在此基础上，我们结晶了两个新的膜蛋白系统，MscS和ABC转运蛋白，实现了分子的跨膜通道。SSRL的高强度晶体学光束线对于天然数据收集、在结构测定中使用MAD定相方法以及鉴定这些蛋白质上的配体结合位点至关重要。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We wish to collect x-ray diffraction data from crystals of membrane proteins that mediate the passage of molecules, electrons and information across the membrane bilayer. We have previously determined the structure of the gated mechanosensitive channel of large conductance (MscL) from Mycobacterium tuberculosis, a prokaryotic channel that functions as an osmotic pressure safety valve (Chang et al. Science 282, 2220 (1998)), and the structure of fumarate reductase (FRD) that catalyzes the final step of anaerobic respiration when fumarate is the terminal electron acceptor (Iverson et al., Science 284, 1961 (1999)). Building upon this foundation, we have crystallized two new membrane protein systems, MscS and an ABC transporter, that achieve the transmembrane passage of molecules. The high intensity crystallography beamlines at SSRL will be essential for native data collection, for the use of MAD phasing methods in the structure determinations, and to identify ligand binding sites on these proteins.