SOLUTION SCATTERING OF PURIFIED GMPA OLIGOMERIC STRUCTURES

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 GmpA是最近发现的一种非同源蛋白，来自不对称分裂的原核生物新月柄杆菌（Caulobacter crescentus），其已被证明是细胞极性建立和维持的关键关键，尽管其定位与复制起点相互作用以维持适当亚细胞结构的多蛋白复合物的能力。对GmpA的一级序列的分析表明，GmpA是一种177个氨基酸的蛋白质，脯氨酸含量超过15%，主要在N-末端区域，高谷氨酸含量导致其pI较低，约为4.0。天然凝胶和尺寸排阻色谱表明，从E.杆菌在纯化的缓冲条件下也存在亚状态的证据，尽管水平低得多。我们建议试点研究GmpA使用溶液的X-射线散射分析的寡聚状态的蛋白质在广泛的条件下。这将有助于理解这种复合物的自组装，探索最大的蛋白质/蛋白质复合物稳定的条件，并研究GmpA与我们已经确定为体内相互作用伙伴的其他蛋白质的相互作用。

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.