STRUCTURAL STUDIES OF ABCG2, HOOKWORM AND S AUREUS PROTEINS

ABCG2、钩虫和金黄色葡萄球菌蛋白质的结构研究

• 批准号: 8361732
• 负责人: OLUWATOYIN Ajibola ASOJO
• 金额: $ 0.55万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361732
• 关键词: ABCG2 gene; ATP phosphohydrolase; ATP-Binding Cassette Transporters; Binding; Biological Assay; Breast; Colon; Data; Development; Drug Transport; Environment; Fibroblasts; Funding; Grant; Hookworms; Institutes; Length; Lung; Malignant Neoplasms; Measures; Mentored Research Scientist Development Award; Mentors; Multi-Drug Resistance; National Center for Research Resources; Nucleotides; Ovary; Pharmaceutical Preparations; Phase; Positioning Attribute; Principal Investigator; Proteins; Research; Research Infrastructure; Resistance; Resources; Roentgen Rays; Source; Staphylococcus aureus; Structure; Testing; Therapeutic; Time; United States National Institutes of Health; anticancer research; cancer cell; chemotherapy; cost; cytotoxicity; mutant; novel; skills; structural biology; three dimensional structure

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. Multidrug resistance is a major obstacle to curing cancer because cancer cells become resistant to diverse and unrelated therapeutic compounds. A mechanism for multidrug resistance is the active extrusion of chemotherapeutic drugs from cancer cells by ABC transporters. ABCG2 is a promiscuous ABC transporter of many unrelated compounds. Mutant forms of ABCG2 are expressed in elevated levels in multidrug resistant cancers of diverse origins including fibroblasts, breast, colon, lung, and ovaries. The mechanisms of drug transport remain unclear and the functions of each domain of ABCG2 are neither tested nor confirmed. Furthermore, there are no 3-dimensional structures of ABCG2. The following specific aims are proposed to rectify this situation: 1) To characterize full length ABCG2 and its domains. The activity of each domain of ABCG2 will be tested using assays that measure cytotoxicity, ATPase activity, drug binding and drug extrusion. 2) To determine 3-dimensional structures of ABCG2's cytosolic domain. X-ray structures will be solved that reveal the mode of binding of nucleotides to the cytosolic domain. 3) To determine 3-dimensional structures of full length ABCG2 and domains. Structures will be solved of full length and active domains of ABCG2. The correlation of structural and activity data will clarify the mechanism of drug transport by ABCG2 and homologous transporters, thus spearheading new strategies for the development of novel chemotherapies for multidrug resistant cancer. Specific aims 1 and 2 are proposed for Phase I, while specific aim 3 is proposed for Phase II of the K01 award and beyond. The K01 will afford the applicant protected time to develop new skills and to apply existing skills to cancer research, with the guidance of her mentors, at the stimulating educational environment of the Eppley Institute. She will then be ready to successfully compete for and obtain an independent tenure track position in cancer research.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 多药耐药是治愈癌症的主要障碍，因为癌细胞对不同和无关的治疗化合物产生抗药性。多药耐药的一个机制是ABC转运蛋白主动将化疗药物从癌细胞中排出。ABCG2是一种混杂的ABC转运蛋白，可以运输许多无关的化合物。ABCG2的突变形式在不同来源的多药耐药癌症中表达水平升高，包括成纤维细胞、乳腺癌、结肠癌、肺癌和卵巢癌。药物转运的机制尚不清楚，ABCG2各个结构域的功能也没有得到测试和证实。此外，ABCG2没有三维结构。为了纠正这种情况，提出了以下具体目标：1)对全长ABCG2及其结构域进行表征。ABCG2的每个区域的活性将通过测量细胞毒性、ATPase活性、药物结合和药物排泄的测试来测试。2)确定ABCG2‘S胞浆结构域的三维结构。将解决揭示核苷酸与胞浆结构域结合模式的X射线结构。3)确定ABCG2全长及其结构域的三维结构。将解决ABCG2的全长和活性结构域的结构。结构和活性数据的关联将阐明ABCG2及其同源转运体的药物转运机制，从而为开发治疗多药耐药癌症的新的化疗药物开辟新的策略。具体目标1和2是为第一阶段提出的，而具体目标3是为K01奖第二阶段及以后提出的。K01将为申请者提供受保护的时间来开发新的技能，并在她的导师的指导下，在Eppley研究所刺激的教育环境中将现有技能应用于癌症研究。然后，她将准备好成功竞争并获得癌症研究领域的独立终身教职。