Anesthetic Bound Calmodulin:Structure Dynamics Function

麻醉结合钙调蛋白：结构动力学函数

• 批准号: 6924288
• 负责人: John H Streiff
• 金额: $ 5.99万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6924288
• 关键词: anesthetics; binding sites; calcium ion; calmodulin; calorimetry; chemical binding; chemical kinetics; chemical structure function; drug interactions; enzyme induction /repression; enzyme inhibitors; enzyme mechanism; fluorescence spectrometry; hydropathy; ligands; myosin light chain kinase; nuclear magnetic resonance spectroscopy; pharmacokinetics; protein sequence; protein structure function; saturated /unsaturated bonds; stop flow technique

DESCRIPTION (provided by applicant): Anesthetic effects on cellular activity manifest as changes in protein function, which might result from direct anesthetic-protein interactions. It is difficult to correlate anesthetic effects on protein function with effects on protein structure and dynamics, in part because there is a dearth of structures of anesthetic-protein complexes. Anesthetics and other hydrophobic drugs are known to affect the ability of calmodulin (CaM) to bind ligands and activate enzymes. The binding of volatile anesthetics to Calmodulin (CaM) is Ca2+- dependent, which indicates they bind to locations that are either not present or not accessible in apoCaM. This makes a compelling argument for determining the structure of an anesthetic-CaM complex, because the structure would reveal the component(s) of Ca2+-CaM structure specific to anesthetic binding. In this project, two specific aims are proposed: A) determine the structure of a volatile anesthetic-CaM complex and B) determine how volatile anesthetics affect CaM function. In specific aim A, NMR methods will be used to estimate the Kd of an anesthetic-CaM complex; determine the structure of an anesthetic-CaM complex, and; determine the effects of anesthetic on the dynamics of CaM. This work will use NMR methods to determine . structure because the Ca2+-CaM resonances have already been assigned, which will expedite the process, and NMR methods are able to elucidate the effects of anesthetics on CaM dynamics. In specific aim B, the effects of anesthetic binding on the Ca2+ affinity of CaM, the binding of peptides to CAM, and the ability of CaM to activate enzymes will be studied with standard biochemical techniques. When this project is completed, it will have demonstrated that volatile anesthetics bind to CaM under specific conditions, determined the solution structure of a volatile anesthetic-CaM complex, and explained how volatile anesthetics might affect the interaction of CaM with proteins.

描述（由申请人提供）： 对细胞活性的麻醉作用表现为蛋白质功能的变化，这可能是由直接麻醉蛋白相互作用引起的。很难将麻醉对蛋白质功能的作用与对蛋白质结构和动力学的影响相关联，部分原因是麻醉蛋白复合物的结构缺乏。已知麻醉药和其他疏水药物会影响钙调蛋白（CAM）结合配体和激活酶的能力。挥发性麻醉与钙调蛋白（CAM）的结合是Ca2+ - 依赖性，这表明它们与apocam中不存在或无法访问的位置结合。这为确定麻醉剂摄像头复合物的结构而引人注目，因为该结构将揭示Ca2+-CAM结构特定于麻醉结合的组件。在该项目中，提出了两个具体的目的：a）确定挥发性麻醉剂摄像头复合物的结构，b）确定挥发性麻醉剂如何影响CAM功能。在特定目的A中，NMR方法将用于估计麻醉胶卷复合物的KD。确定麻醉胶卷复合物的结构，并；确定麻醉对CAM动力学的影响。这项工作将使用NMR方法来确定。结构是因为已经分配了CA2+-CAM共振，这将加快该过程，并且NMR方法能够阐明麻醉药对CAM动力学的影响。在特定的目标B中，麻醉结合对CAM的Ca2+亲和力的影响，肽与CAM的结合以及CAM激活酶的能力将使用标准的生化技术研究。该项目完成后，它将证明挥发性麻醉药在特定条件下与CAM结合，确定挥发性麻醉剂胶体复合物的溶液结构，并解释挥发性麻醉药如何影响CAM与蛋白质与蛋白质的相互作用。