MEMBRANE TRANSPORTERS

膜转运蛋白

• 批准号: 7957239
• 负责人: Christopher Miller
• 金额: $ 1.75万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957239
• 关键词: Acids; Agmatine; Arginine; Computer Retrieval of Information on Scientific Projects Database; Crystallography; Decarboxylation; Escherichia coli; Family; Funding; Grant; Home environment; Institution; L-Selenomethionine; Light; Membrane; Membrane Proteins; Phase; Proteins; Proton Pump; Research; Research Personnel; Resistance; Resources; Source; Synchrotrons; United States National Institutes of Health; improved; indexing; overexpression; protein E; response; virtual

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. This is a "MFS-family" membrane protein from E coli that stoichiometircally exchanges arginine for its decarboxylation product, agmatine. It operates as a "virtual proton pump" in the extreme acid resistance response. This protein can be easily overexpressed and crystallized, and we are currently trying to improve crystal quality to the point where we can begin serious indexing and phasing. During the past year, we have improved diffraction limit from about 20 A to 6A in the best, isotropic cases. Our trips to NSLS are used for screenng and optimization at this point. This cannot be done on our home source, as these membrane protein crystals require a high-intensity beam to assess their diffration limit. If we can achieve isotropic diffraction out to 3.5 A, we will then begin phasing, initially using MAD with SeMet derivatives, which we can readily express.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 这是一种来自e大肠杆菌的“ MFS家族”膜蛋白，将精氨酸的脱羧产物agmatine换成精氨酸。 它在极高的酸性响应中充当“虚拟质子泵”。 这种蛋白质很容易过表达和结晶，我们目前正在尝试提高晶体质量，以至于我们可以开始认真的索引和相位。在过去的一年中，在最佳的各向同性病例中，我们的衍射极限从约20 a增加到6A。 此时，我们对NSL的旅行用于ScreenNG和优化。 这不能在我们的家用来源上完成，因为这些膜蛋白晶体需要高强度光束来评估其衍射极限。 如果我们可以将各向同性的衍射量超出3.5 a，那么我们将开始使用SEMET衍生物MAD，我们可以很容易地表达。