INTEGRAL MEMBRANE PROTEINS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 整合膜蛋白占蛋白质组的约30%，并且在代谢、调节和细胞间过程中起关键作用。 人类MP是所有治疗药物的约40%的靶标，但MP结构的数量小于可溶性蛋白质3D结构数量的0.5%。 在我们的实验室，作为膜蛋白结构中心（CSMP）和膜蛋白表达中心（MPEC）的成员，我们的目标是确定具有高生物医学影响的整体MP结构。 我们的中心努力通过提供来自E.大肠杆菌、极端微生物、酵母菌和人类。 我们的表达能力包括原核和真核（包括HEK）体内系统，以及E。基于大肠杆菌的无细胞体外系统优化MP表达。 蛋白质纯化核心实验室包括表征方法，并提供纯净均匀和稳定的蛋白质，不含过量的洗涤剂。 包括X射线衍射的结构测定是由机器人晶体试验和衍射在先进的光源光束线8.3.1，世界上最具生产力的蛋白质晶体学设施之一的辅助。 质谱分析将有助于确认每种蛋白质的质量和均一性，因为一些膜蛋白具有切割的N-末端或进行翻译后修饰。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Integral membrane proteins account for ~30% of a proteome and play critical roles in metabolic, regulatory and intercellular processes. Human MPs are the targets for ~40% of all therapeutic drugs, but the number of MP structures is less than 0.5% of the number of soluble protein 3D structures. In our lab and as members of the Center for Structure of Membrane Proteins (CSMP) and Membrane Protein Expression Center (MPEC), we aim to determine integral MP structures of high biomedical impact. Our centers strive to obtain structures by providing many targets from E. coli, extremophiles, yeast and humans. Our expression capabilities include prokaryotic and eukaryotic (including HEKs) in vivo systems, and an E. coli based cell-free in vitro system optimized for MP expression. The protein purification core lab includes characterization methods and provides pure homogeneous and stable proteins free of excess detergent. Structure determination by include X-ray diffraction is aided by robotic crystal trials and diffraction at the Advanced Light Source beamline 8.3.1, one of the world's most productive protein crystallography facilities. Mass Spectroscopy analysis will be useful in confirming each protein mass and homogeneity, since some membrane protein have cleaved N-termini or are subject to post-translational modification.