OLIGOMERIC STATE OF VPS4: HIV

VPS4 的寡聚状态：HIV

• 批准号: 7598200
• 负责人: CHRISTOPHER P. HILL
• 金额: $ 0.23万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598200
• 关键词: ATP phosphohydrolase; Binding; Cells; Computer Retrieval of Information on Scientific Projects Database; Condition; Enzymes; Funding; Grant; HIV; Institution; Modeling; Pathway interactions; Preparation; Proteins; Publishing; Recruitment Activity; Research; Research Personnel; Resources; Sampling; Shapes; Solutions; Source; Structure; Testing; Time; United States National Institutes of Health; Vacuolar Protein Sorting; Virion; Virus; day; molecular mass; molecular shape; nucleotide analog; protein function; size

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. The Vps4 protein is the AAA ATPase that drives progression through the vacuolar protein sorting pathway. This pathway delivers substrates and resident enzymes to lysozome precursors. HIV and other enveloped viruses recruit this pathway to bud virus particles from host cells. We recently published the structure of Vps4 in a monomeric state (EMBO J. 24, 3658-69, 2005). Because the protein functions as an oligomer, we now seek to test models for the assembled state. Leading models include 12-mer, 10-mer, 6-mer, or 5-mer, with either single or double disk configurations. We routinely prepare highly pure samples that are soluble to at least 20 mg/ml. Preparations take just a few days to complete. Under appropriate conditions, such as bound nucleotide analog, the protein elutes from a sizing column as a well defined oligomer. The retention time corresponds to a ~9-11 mer, but of course, retention time is highly dependent upon molecular shape, and we seek better estimates of solution molecular mass and shape.

这个子项目是许多研究子项目中的一个 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， 因此可以在其他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。 Vps 4蛋白是AAA ATP酶，通过液泡蛋白分选途径驱动进展。 该途径将底物和驻留酶递送至溶菌酶前体。 HIV和其他包膜病毒通过这一途径从宿主细胞中出芽。 我们最近公开了单体状态的Vps 4的结构（EMBO J.24，3658-69，2005）。 由于蛋白质的功能是作为一个寡聚体，我们现在试图测试组装状态的模型。 主要模型包括12聚体、10聚体、6聚体或5聚体，具有单盘或双盘配置。我们常规制备可溶性至少为20 mg/ml的高纯样品。 准备工作只需几天即可完成。 在适当的条件下，例如结合的核苷酸类似物，蛋白质从大小测定柱中洗脱为明确定义的寡聚体。 保留时间对应于~9-11聚体，但当然，保留时间高度依赖于分子形状，并且我们寻求更好地估计溶液分子质量和形状。