Core C: Protein Chemistry Core

核心 C：蛋白质化学核心

• 批准号: 7777114
• 负责人: JONATHAN Brewer COHEN
• 金额: $ 28.52万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777114
• 关键词: Acetylcholine; Amino Acids; Anesthetics; Binding; Binding Sites; Biochemical; Blood capillaries; Brain; Chemical Structure; Core Facility; Drug Binding Site; Equipment; Fluorescence; Gated Ion Channel; General anesthetic drugs; Instruction; Integral Membrane Protein; Laboratories; Ligands; Location; Mass Spectrum Analysis; Modeling; Molecular Models; Peptides; Protein Chemistry; Proteins; Radioactive; Research; Research Personnel; Research Project Grants; Resources; Sampling; Sequence Analysis; Serotonin; Serotonin Receptors 5-HT-3; Site; Structure; Supervision; System; Technical Expertise; Techniques; capillary; citrate carrier; computing resources; design; detector; molecular modeling; programs; receptor

CORE C: Protein Chemistry Core The Protein Chemistry Core will continue to provide for the research groups in the Program Project the technical expertise and equipment required for the isolation and sequence analysis of receptor fragments containing the sites of photoincorporation by general anesthetics. In addition, the Core provides computational resources for the molecular modeling that is necessary to interpret the protein chemistry results in terms of models of receptor structure and to provide guidance in the design of mutational analyses necessary to define the energetic contributions to anesthetic binding. The Core is under the supervision of Dr. Cohen and is located in his laboratory. Major equipment cunrently available in the Core include (i) an Applied Biosystems 492 Precise automated Protein Sequenator and in-line amino acid analyzer; and (ii) 2 Agilent 1100 HPLCs equipped with UV and fluorescence detectors; and (iii) an Agilent 1100 capillary LC system. The biochemical characterization of anesthetic binding sites in ligand-gated ion channels requires the use of highly specialized techniques not normally available in protein chemistry core facilities which usually do not accept radioactive samples either for Edman degradation or mass spectrometry. The identification of drug binding sites within the hydrophobic domains of integral membrane proteins poses unique problems for peptide isolation and characterization by either Edman degradation or mass spectrometry. It is the function of the Core to provide the appropriate equipment and highly skilled protein chemists who can interact directly with the research projects to develop and carry out the appropriate research strategies and to educate the investigators about the necessary protein chemistry techniques to be carried out either in the Core or in the investigator's lab. RELEVANCE (See instructions): GABAARS in the brain are a major site of action of many clinically used anesthetics of diverse chemical structure, but the number and location of anesthetic binding sites within GABAARS or in the structurally related nicotinic acetylcholine and serotonin 5-HT3 receptors remains unknown. This Core provides the resources necessary to identify these binding sites.

核心C：蛋白质化学核心 蛋白质化学核心将继续为计划项目中的研究小组提供 隔离和序列分析受体片段所需的技术专长和设备 包含通用麻醉药的光企业部位。此外，核心提供 解释蛋白质化学所必需的分子建模的计算资源 根据受体结构模型的结果，并在突变分析的设计中提供指导 定义对麻醉结合的能量贡献所必需的。核心在 科恩博士，位于他的实验室。核心中可用的主要设备包括（i） Applied Biosystems 492精确的自动化蛋白序列机和在线氨基酸分析仪； （ii）2 配备紫外线和荧光探测器的安捷伦1100 HPLC； （iii）一个敏捷的1100毛细管LC 系统。配体门控离子通道中麻醉结合位点的生化表征需要 蛋白质化学核心设施中通常不可用的高度专业技术的使用 通常不接受Edman降解或质谱的放射性样品。这 鉴定整体膜蛋白疏水结构域中的药物结合位点构成 Edman降解或质量的肽隔离和表征的独特问题 光谱法。提供适当的设备和高技巧的蛋白质是核心的功能 可以直接与研究项目互动以开发和执行适当的化学家 研究策略并教育研究人员有关必要的蛋白质化学技术 在核心或调查员实验室中进行。 相关性（请参阅说明）： 大脑中的Gabaars是许多临床使用的多种化学药品麻醉药的主要作用部位 结构，但在加巴人或结构上的麻醉结合位点的数量和位置 相关的烟碱乙酰胆碱和5-羟色胺5-HT3受体仍然未知。这个核心提供了 确定这些约束地点所需的资源。