ELECTRON MICROSCOPY OF MEMBRANE PROTEINS(RMI)

膜蛋白电子显微镜 (RMI)

• 批准号: 7262975
• 负责人: RONALD A MILLIGAN
• 金额: $ 31.38万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-23 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7262975
• 关键词: ATP-Binding Cassette Transporters; Accounting; Binding; Cells; Communication; Condition; Cryoelectron Microscopy; Crystallization; Data; Detergents; Development; Electron Microscopy; Exhibits; Hand; Helium; Helix (Snails); Image; Image Analysis; Investigation; Laboratories; Maps; Mediating; Medical; Membrane; Membrane Proteins; Membrane Transport Proteins; Microscope; Multi Antimicrobial Toxin Extrusion; Nucleotides; Numbers; Organelles; Organism; Process; Proteins; Range; Research; Research Personnel; Resistance; Resolution; Risk; Screening procedure; Speed; Staining method; Stains; Structure; Suggestion; System; Transport Process; Tube; Work; antineoplastic antibiotics; ear helix; experience; improved; insight; member; research study; success

DESCRIPTION (provided by applicant): Membrane proteins account for approximately 25% of gene products in organisms. They populate cell and organelle membranes and are essential for communication across these hydrophobic compartmental boundaries. They participate in recognition, transport and transduction processes that are fundamental attributes of living systems. In the work described here, the structure and mechanism of action of a number of membrane protein transporters will be investigated by cryo-electron microscopy and image analysis. Transporters from the ATP Binding Cassette (ABC), Major Facilitator (MFS) and Multi Antimicrobial Toxin Extrusion (MATE) superfamilies will be studied, as these are the current focus of parallel x-ray crystallographic investigations by one of the investigators. Members of the ABC, MFS and MATE superfamilies are of great medical importance as they are responsible for antibiotic and chemotherapy resistance. Cryo-EM and image analysis will be used to calculate 3D maps from (i) helical tubes and 2D crystals of MsbA - an ABC transporter - in various nucleotide-bound and substrate-bound states and, (ii) 2D crystals of FucP - an MFS transporter - with substrate bound. The maps will reveal the structures and provide insights into the transport mechanisms of representative members of these two membrane protein superfamilies. Additional experiments will focus on screening crystallization conditions to improve the order of the existing helical and 2D arrays, and to grow ordered arrays of other membrane proteins for EM structural studies.

描述（由申请人提供）：膜蛋白约占生物体中基因产物的25%。它们分布在细胞和细胞器膜上，对跨越这些疏水隔室边界的通信至关重要。它们参与识别、运输和转导过程，这些都是生命系统的基本属性。