NANOASSAY FOR REALTIME MOLECULAR PROBING ABC TRANSPORTER

用于实时分子探测 ABC 转运蛋白的纳米测定

• 批准号: 8361112
• 负责人: X. Nancy Xu
• 金额: $ 1.23万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361112
• 关键词: ATP Hydrolysis; ATP-Binding Cassette Transporters; Antibiotic Resistance; Bacteria; Binding; Biological; Biomedical Research; Complex; Computers; Coupling; Cystic Fibrosis; Development; Electron Microscopy; Family; Functional disorder; Funding; Fungal Drug Resistance; Grant; Image; Ions; Life; Lipids; Mechanics; Membrane Proteins; Mining; Molecular; Molecular Probes; Molecular Structure; National Center for Research Resources; Nucleotides; Organism; Peptides; Principal Investigator; Pump; Research; Research Infrastructure; Resistance; Resources; Source; Structure; Transmembrane Domain; Transmission Electron Microscopy; United States National Institutes of Health; base; cancer cell; cost; macromolecule; member; nanoassay; reconstruction; structural biology; sugar; uptake

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. ABC transporters are one of the largest and most diverse super-families of membrane proteins found in all living organisms ranging from bacteria to humans.1-5 These membrane proteins act as mechanical pumps that couple ATP hydrolysis to either uptake or export of a wide diversity of substrates, including ions, sugars, lipids, peptides or complex organic molecules, against concentration gradients, across biological membranes.1-5 Interestingly, all ABC transporters share a common organization of a four core domains: two transmembrane domains (TMD) and two nucleotide-binding domains (NBD), indicating a common evolutionary origin and suggesting a similar mechanism of energy coupling. ABC transporters are medically relevant because they can be responsible for antibiotic and antifungal resistances in micro-organisms, resistance to chemotherapeutic treatments in cancer cells, and dysfunctions of many eukaryotic members are the molecular basis for severe sicknesses (e.g., cystic fibrosis).5-7 Unfortunately, despite extensive studies, many questions about the molecular mechanism and interactions of ABC transporters remain unanswered. Built upon the significant progress that we have made and the recent development of cryo-transmission electron microscopy (cryo-TEM), in this revised proposal, we aim to expand our original proposed research scope to use cryo-TEM, computer reconstruction and structure mining to solve the structure and molecular mechanism of BmrA, a multidrug bacterial transporter belonging to the ABC transporter superfamily, by collaborating with the National Center for Macromolecular Imaging (NCMI) (a NCRR designated Biomedical Research Resource for Structural Biology, at http://ncmi.bcm.tmc.edu/).

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 ABC 转运蛋白是从细菌到人类的所有生物体中发现的最大、最多样化的膜蛋白超家族之一。1-5 这些膜 蛋白质充当机械泵，将 ATP 水解与多种底物的吸收或输出耦合起来，包括离子、糖、脂质、肽或复杂的有机物 分子，针对浓度梯度，跨生物膜。1-5 有趣的是，所有 ABC 转运蛋白都具有四个核心结构域的共同组织：两个跨膜 结构域（TMD）和两个核苷酸结合结构域（NBD），表明共同的进化起源并表明类似的能量耦合机制。 ABC 转运蛋白具有医学相关性，因为它们可能导致微生物对抗生素和抗真菌药物产生耐药性，以及对癌症化疗产生耐药性 细胞和许多真核成员的功能障碍是严重疾病（例如囊性纤维化）的分子基础。5-7 不幸的是，尽管进行了广泛的研究，但关于 ABC 转运蛋白的分子机制和相互作用仍未得到解答。 基于我们取得的重大进展和冷冻透射电子显微镜 (cryo-TEM) 的最新发展，在本修订提案中，我们的目标是扩大我们的研究范围 最初提出的研究范围是利用冷冻透射电镜、计算机重建和结构挖掘来解决多药细菌BmrA的结构和分子机制 属于 ABC 转运蛋白超家族的转运蛋白，通过与国家大分子成像中心 (NCMI)（NCRR 指定的生物医学研究资源中心）合作 结构生物学，网址：http://ncmi.bcm.tmc.edu/）。