STRUCTURAL AND FUNCTIONAL STUDIES OF P97

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们正在进行p97（也称为VCP，含缬素蛋白；在古细菌中称为VAT；在酵母中称为cdc48）的结构和功能研究，它是NSF的远缘同源物。p97与有丝分裂后内质网（ER）和高尔基体的同型膜融合、细胞核的组装和生长、细胞凋亡和DNA复制有关。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进行细胞质降解。我们最近确定了生物完整的p97/VCP六聚体与ADP和ADP混合物络合的晶体结构。AlFx使用在SSRL和APS收集的衍射数据。在SSRL收集的MAD数据集提供了解决结构的关键。该结构揭示了单个结构域和连接连接体的第四纪构型。我们的晶体结构代表了p97/VCP在其核苷酸水解循环过程中的快照。我们的未来计划包括对p97/VCP的连接蛋白和底物的复合物进行结构研究，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.