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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 大约有80种不同的钾通道，这对于活细胞至关重要。 这些被分类为电压门控，内部整流，依赖性钙和核。 KCSA是一种细菌配体门控钾通道，可通过细胞内质子打开刺激。 对于WT蛋白和众多突变体，KCSA在各种条件下都成功地结晶了。 另一方面，Shaker Channe是一个电压门控的K+通道，其结构与KCSA相似，但Shaker对结晶提出了更加困难的挑战。因此，在一项副本中，我们的目的是通过构建各种KCSA-shaker嵌合蛋白来研究振荡器的结构和功能方面。 外膜蛋白F（OMPF）是允许抗生素分子渗透到革兰氏阴性细菌细胞的主要门户。 随着膜渗透性降低的出现，抗生素耐药性的出现，有必要找到增加OMPF PORIN抗生素易位的方法。 因此，在第二个副标中，我们与各种抗生素中的复合物中结晶了OMPF。 我们第一次能够在分子水平上实验可视化蛋白质 - 药物相互作用。 因此，这项工作的结果将洞悉具有改善扩散特征和目标特异性的抗生素设计。 介导的离子型谷氨酸受体离子通道（iGlurs）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 这些受体的配体结合域（LBD）驱动开口 跨膜孔。 这些受体作为四聚体组装。 但是，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。