SOLUTION SCATTERING OF PURIFIED GMPA OLIGOMERIC STRUCTURES

纯化 GMPA 寡聚结构的溶液散射

• 批准号: 7954469
• 负责人: Thomas N Earnest
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954469
• 关键词: Amino Acids; Architecture; Buffers; Caulobacter crescentus; Cell Polarity; Complex; Computer Retrieval of Information on Scientific Projects Database; Escherichia coli; Funding; Gel; Glutamic Acid; Grant; Homologous Protein; Institution; Maintenance; Molecular Sieve Chromatography; N-terminal; Pilot Projects; Proline; Proteins; Replication Origin; Research; Research Personnel; Resources; Solutions; Source; Structure; United States National Institutes of Health; in vivo; overexpression; protein complex; self assembly; structural biology; synchrotron radiation

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. GmpA is a recently discovered non-homologous protein from the assymetrically-dividing procaryote, Caulobacter crescentus, which has been shown to be a critical lynchpin in the establishment and maintenance of cell polarity though its ability to a localize a multi-protein complex which interacts with the origin of replication to maintain the proper subcellular architecture. Analysis of the primary sequence of GmpA shows a 177 amino acid protein with a proline content of over 15%, primarily in the N-terminal region, and a high glutamic acid content contributing to its low pI of ~4.0. Native gels and size exclusion chromatography indicate a higher-ordered oliomeric state when purified from overexpression in E. coli. Evidence of substates under the buffer conditions of purification is also present, though at a much lower level. We propose pilot studies of GmpA using solution x-ray scattering to analyse the oligomeric state of the protein under a wide range of conditions. This will assist in understanding the self-assembly of this complex, explore conditions of maximal protein/protein complex stabilization, and study interactions of GmpA with other proteins that we have identified as interaction partners in vivo.

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。 GMPA是一种最近发现的非同源蛋白质，来自分解的procaryote caulobacter crescentus，它已被证明是细胞极性建立和维持细胞极性的关键lynchpin，尽管其能够与多蛋白复合物的本地化能力相互作用，从而与复制的起源相互作用，以维持合适的亚纤维构造。对GMPA的一级序列的分析显示，脯氨酸含量超过15％，主要在N末端区域，以及高谷氨酸含量，其低PI含量为〜4.0。天然凝胶和大小排除色谱法表明当从大肠杆菌中的过表达纯化时，纯度的橄榄液状态。在纯化的缓冲条件下，在较低的水平上也存在纯化的证据。我们建议使用溶液X射线散射对GMPA进行试验研究，以在广泛的条件下分析蛋白质的寡聚状态。这将有助于理解这种复合物的自组装，探索最大蛋白/蛋白质复合物稳定的条件，并研究GMPA与我们已确定为体内相互作用伙伴的其他蛋白质的相互作用。