STRUCTURAL AND FUNCTIONAL STUDIES OF P97

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。我们正在进行p97(也被称为VCP，含有Valosin的蛋白质；在古细菌中的VAT；在酵母中的CDC48)的结构和功能研究，它是NSF的远程同源物。P97参与了内质网和高尔基体的有丝分裂后同型膜融合、细胞核的组装和生长、细胞凋亡和DNA复制。它还参与泛素依赖的降解途径，以及从内质网中提取错误折叠的管腔和膜蛋白用于胞浆降解。我们最近利用在SSRL和APS收集的衍射数据确定了与ADP和ADP.AlFx的混合物形成的生物完整的p97/VCP六聚体的晶体结构。SSRL收集的MAD数据集为解决这一结构提供了关键。该结构揭示了单个结构域和连接连接子的四元构型。我们的晶体结构代表了p97/VCP在核苷酸水解循环中的快照。我们未来的计划包括研究p97/VCP的接头蛋白和底物的复合体的结构，以阐明p97/VCP的机制和生物学作用。

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. We are pursuing structural and functional studies of p97 (also called VCP, valosin-containing protein; VAT in Archaebacteria; and cdc48 in yeast), a distant homolog of NSF. p97 has been implicated in postmitotic homotypic membrane fusion of the endoplasmic reticulum (ER) and of the Golgi apparatus, assembly and growth of the nucleus, apoptosis, and DNA replication. It is also involved in the ubiquitin-dependent degradation pathway, as well as the extraction of misfolded luminal and membrane proteins from the ER for cytosolic degradation. We recently determined the crystal structure of the biologically complete p97/VCP hexamer complexed with a mixture of ADP and ADP.AlFx using diffraction data collected at SSRL and APS. The MAD dataset collected at SSRL provided the key to solve the structure. The structure revealed the quarternary configuration of the individual domains and the connecting linkers. Our crystal structure represents a snapshot of p97/VCP as it proceeds through its nucleotide hydrolysis cycle. Our future plans include structural studies of complexes involving adapter proteins and substrates for p97/VCP, to elucidate the mechanism and biological role of p97/VCP.