Opioid Receptor Protein Production Core

阿片受体蛋白生产核心

• 批准号: 9054101
• 负责人: Vadim Cherezov
• 金额: $ 32.38万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9054101
• 关键词: ADORA2A gene; Adverse effects; Agonist; Biomass; Collaborations; Complex; Crystallization; Cytochromes; G-Protein-Coupled Receptors; Generations; Instruction; Lead; Libraries; Ligand Binding; Ligands; Maintenance; Monitor; Nature; Nociception; Opioid Receptor; Peptide Receptor; Preparation; Production; Proteins; Protocols documentation; Quality Control; Resolution; Sampling; Serotonin Receptor Binding; Structure; Work; design; drug discovery; mutant; programs; receptor; research study; therapeutic target

Production of large quantities of hiighly purified receptors is an underlying requirement for all the projects in ttie Program Project -"Structure-Function of Opioid Receptors for Drug Discovery. The production of these samples will be carried out using well established protocols that have now been optimized for use with GPCRs as part of GPCR Network's GPCR Structure Determination Pipeline. These production protocols now include the use of newly developed GPCR fusion partner toolchest for stabilization and crystallization that has increased the quantity and quality of structures that we are able to determine. Most notably, with the use of a new fusion partner, apo-cytochrome b562 (RIL) mutant, BRIL, we were able to determine the structure of the A2A adenosine receptor- ZM241385 to 1.8 A resolution, the structure of NOP, and more recently 2 agonist bound serotonin receptors in an activated state. Structural studies will demand the use of large volumes of highly purified protein for crystallization studies while others doing functional studies will need much less. Studies that led to the structure solution of the K-opioid receptor and the nociception/oprhanin FQ peptide receptor required a large number of constructs and an average of 25 mgs of highly purified protein or about 100 liters of biomass. For receptor-ligand complex structural studies we estimate that 5-10 mgs will be required per structure.

生产大量纯化的受体是所有项目的基本要求 TTIE计划项目 - “阿片类药物受体的结构功能。 样本将使用现已优化供与其一起使用的良好协议进行 GPCR作为GPCR网络GPCR结构确定管道的一部分。这些生产方案 现在包括使用新开发的GPCR融合伙伴工具琴进行稳定和结晶 这增加了我们能够确定的结构的数量和质量。最值得注意的是， 使用新的融合合作伙伴Apo-cytochrome B562（RIL）突变体，Bril，我们能够确定 A2A腺苷受体的结构 - ZM241385至1.8 A分辨率，NOP的结构等等 最近，2种激活状态下的激动剂结合的5-羟色胺受体。结构研究将要求使用 大量高度纯化的蛋白质用于结晶研究，而其他进行功能研究的蛋白质将 需要少得多。导致K-阿片类受体和溶液的研究 伤害感受/oprhanin FQ肽受体需要大量构建体，平均25 mg 高度纯化的蛋白质或约100升生物量。对于受体配体复杂的结构研究，我们 估计每个结构将需要5-10毫克。