STRUCTURAL STUDIES OF AIDS-RELATED ENZYMES AND OTHER PATHOGENIC TARGETS

艾滋病相关酶和其他致病靶标的结构研究

• 批准号: 8362410
• 负责人: Vivian Cody
• 金额: $ 0.1万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362410
• 关键词: Acquired Immunodeficiency Syndrome; Area; Biological; Complex; Data; Dihydrofolate Reductase; Dihydrofolate Reductase Inhibitor; Dihydropteroate Synthase; Dimerization; Drug resistance; Enzymes; Escherichia coli; Funding; Goals; Grant; Human; Immunocompromised Host; Investigation; Medical Research; Minnesota; Mutation; Nanotubes; National Center for Research Resources; Opportunistic Infections; Patients; Pneumocystis; Pneumocystis carinii Pneumonia; Principal Investigator; Radiation; Request for Proposals; Research; Research Infrastructure; Research Institute; Resolution; Resources; Role; Site; Site-Directed Mutagenesis; Source; Structure; Therapeutic Agents; Time; United States National Institutes of Health; Variant; cost; design; dimer; inhibitor/antagonist; mortality; novel; pathogen; programs; structural biology; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Cody lab at the Hauptman-Woodward Medical Research Institute (HWI) has a multi-faceted research program that focuses on understanding the role of site specific residues in the design of selective inhibitors of pathogens such as Pneumocystis (Pc), a major cause of opportunistic infection and mortality in immunocompromised patients, particularly those with AIDS. Pneumocystis jirovecii (pj) is the causative agent of Pneumocystis pneumonia (PcP), one of the most frequent and severe opportunistic infections in immunocompromised patients. In this proposal, we request beamtime to develop complementary areas of biological structural research. Recent studies show that mutations accumulate over time in the target enzymes, dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) from Pneumocystis, potentially giving rise to drug resistance. A major goal of this project is to characterize pjDHFR and pjDHPS and their variants in order to design effective inhibitors that have potential as therapeutic agents for the treatment of PcP. Site-directed mutagenesis studies on DHFR enzymes are under investigation to determine the role of specific residues in modulating DHFR inhibitor potency and in conferring drug-resistance to pjDHFR as observed in AIDS patient isolates. Structures of human DHFR inhibitor complexes are under investigation to compare their structures with those of the Pneumocystis DHFR enzymes. We require the use of synchrotron radiation to determine high-resolution details of these inhibitor complexes. In another collaborative study (Wagner, Univ. Minnesota), E. coli DHFR dimerization complexes are being investigated to design novel nanotube assemblies. Structural data are needed to validate dimerization assembly and to determine how mutations at the dimer interface modulate nanotube assembly. Sychrotron time is needed as these crystals tend to be small and diffract modestly.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 豪普特曼-伍德沃德医学研究所（HWI）的科迪实验室拥有一个多方面的研究计划，重点是了解位点特异性残基在设计肺孢子虫（Pc）等病原体选择性抑制剂中的作用，肺孢子虫是免疫功能低下患者，特别是艾滋病患者机会性感染和死亡的主要原因。耶氏肺孢子虫（Pneumocystis jirovecii，pj）是肺孢子虫肺炎（Pneumocystis pneumonia，PcP）的病原体，是免疫功能低下患者最常见和最严重的机会性感染之一。在这个建议中，我们要求beamtime开发生物结构研究的补充领域。最近的研究表明，随着时间的推移，突变在肺孢子虫的靶酶，二氢叶酸还原酶（DHFR）和二氢蝶酸合酶（DHPS）中积累，可能导致耐药性。该项目的一个主要目标是表征pjDHFR和pjDHPS及其变体，以设计有效的抑制剂，其具有作为治疗PcP的治疗剂的潜力。对DHFR酶的定点诱变研究正在调查中，以确定特定残基在调节DHFR抑制剂效力和赋予艾滋病患者分离株中观察到的pjDHFR耐药性中的作用。人DHFR抑制剂复合物的结构正在研究中，以将其结构与肺孢子虫DHFR酶的结构进行比较。我们需要使用同步辐射，以确定这些抑制剂复合物的高分辨率的细节。在另一项合作研究中（瓦格纳，明尼苏达大学），E.大肠杆菌DHFR二聚复合物正在研究设计新的纳米管组件。需要结构数据来验证二聚组装和确定二聚体界面处的突变如何调节纳米管组装。同步加速时间是必要的，因为这些晶体往往是小的和适度的行为。