STRUCTURE DETERMINATION OF ION CHANNEL PROTEINS

离子通道蛋白的结构测定

• 批准号: 8169300
• 负责人: BENOIT ROUX
• 金额: $ 1.05万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169300
• 关键词: Antibiotic Resistance; Antibiotics; Architecture; Binding; Brain; Calcium; Cell Membrane Permeability; Cells; Characteristics; Chimeric Proteins; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Drug Interactions; Funding; Glutamate Receptor; Grant; Hand; Institution; Ion Channel; Ion Channel Protein; Life; Ligand Binding Domain; Ligands; Mediating; Molecular; Neurotransmitters; OprF protein; Potassium Channel; Proteins; Protons; Research; Research Personnel; Resources; Source; Specificity; Spinal Cord; Stimulus; Structure; Synapses; Time; United States National Institutes of Health; Voltage-Gated Potassium Channel; Work; antibiotic design; dimer; improved; insight; mutant; novel; porin; receptor; response; voltage

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. There are about 80 different potassium channels, which are essential for living cells. These are classified as voltage-gated, inward rectifying, Calcium dependent, and prokaryotic. KcsA is a bacterial ligand gated potassium channel that opens upon stimulus by intracellular protons. KcsA has been successfully crystallized in various conditions for both the WT protein and for numerous mutants. Shaker channel, on the other hand, is a voltage gated K+ channel with similar architecture to KcsA, but Shaker presents a more difficult challenge for crystallization. Therefore, in one subproject, our aim is to study the structural and functional aspects of Shaker by constructing various KcsA-Shaker chimera proteins. Outer membrane protein F (OmpF) is the main gateway that allows antibiotic molecules to permeate Gram-negative bacterial cells. With the emergence of antibiotic resistance by virtue of decreased membrane permeability, it is necessary to find ways to increase antibiotic translocation across OmpF porin. Therefore, in a second subproject, we have crystallized OmpF in complex with various antibiotics. For the first time, we are able to experimentally visualize protein-drug interactions at the molecular level. Therefore, results of this work will give insights into the design of antibiotics with improved diffusional characteristics and target specificity. The ionotropic glutamate receptor ion channels (iGluRs) mediate excitatory responses at the vast majority of synapses in the brain and spinal cord. The binding of neurotransmitter molecules to the ligand-binding domain (LBD) of these receptors drives the opening of the transmembrane pore. These receptors assemble as tetramers. However, previous crystal structures have revealed only dimer complexes. Therefore, in this third project, we have solved the crystal structure of a novel GluR2 LBD dimer-of-dimers.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 大约有 80 种不同的钾通道，对活细胞至关重要。 这些被分类为电压门控型、内向整流型、钙依赖性型和原核型。 KcsA 是一种细菌配体门控钾通道，在细胞内质子刺激下打开。 KcsA 已在各种条件下成功结晶为 WT 蛋白和众多突变体。 另一方面，Shaker 通道是电压门控 K+ 通道，其架构与 KcsA 类似，但 Shaker 对结晶提出了更困难的挑战。因此，在一个子项目中，我们的目标是通过构建各种 KcsA-Shaker 嵌合蛋白来研究 Shaker 的结构和功能。 外膜蛋白 F (OmpF) 是抗生素分子渗透革兰氏阴性细菌细胞的主要通道。 随着膜通透性降低而出现抗生素耐药性，有必要找到增加抗生素跨 OmpF 孔蛋白易位的方法。 因此，在第二个子项目中，我们将 OmpF 与各种抗生素复合结晶。 我们第一次能够在分子水平上通过实验可视化蛋白质-药物相互作用。 因此，这项工作的结果将为设计具有改善的扩散特性和靶点特异性的抗生素提供见解。 离子型谷氨酸受体离子通道 (iGluR) 介导 大脑中绝大多数突触的兴奋反应 脊髓。神经递质分子与神经递质的结合 这些受体的配体结合域（LBD）驱动 跨膜孔。 这些受体组装成四聚体。 然而，以前的晶体结构仅揭示了二聚体复合物。 因此，在第三个项目中，我们解决了新型 GluR2 LBD 二聚体的晶体结构。