Structural Characterization of Gbetagamma-Phospholipase C complexes

Gbetagamma-磷脂酶 C 复合物的结构表征

• 批准号: 10551201
• 负责人: ISAAC FISHER
• 金额: $ 7.42万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10551201
• 关键词: Absence of pain sensation; Address; Analgesics; Binding; Binding Proteins; Binding Sites; Biological Assay; C-terminal; Cardiovascular Diseases; Cardiovascular system; Cell membrane; Cells; Chemicals; Complex; Complex Analysis; Consensus; Cryoelectron Microscopy; Data; Deuterium; Development; Disease; Distal; EF Hand Motifs; Enzymes; Family; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Health; Healthcare; Heart Diseases; Heart Hypertrophy; Heterotrimeric G Protein Subunit; Hydrogen; In Vitro; Inflammation; Learning; Lipase; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane; Molecular; Molecular Conformation; Mutagenesis; Opioid; Opioid Analgesics; PH Domain; Pathology; Pathway interactions; Phosphatidylinositol 4,5-Diphosphate; Phospholipase; Phospholipase C; Physiology; Pre-Clinical Model; Process; Protein Isoforms; Protein Kinase C; Proteins; Regulation; Research; Resolution; Roentgen Rays; Role; Second Messenger Systems; Signal Transduction; Site-Directed Mutagenesis; Structure; Sulfhydryl Compounds; Surface; Testing; Therapeutic; United States; crosslink; improved outcome; insight; mutant; non-opioid analgesic; novel; opioid epidemic; particle; prevent; protein protein interaction; protein structure; reconstruction; response; tool; treatment strategy

PROJECT SUMMARY The opioid crisis, cardiovascular disease, and cancer are the most pressing healthcare crises in the United States. PLCβ enzymes are essential for normal analgesic, cardiovascular, and proliferative signaling. Further, disruption of interactions between PLCβ and its G protein activators have been shown to improve outcomes in preclinical models of opioid induced analgesia, cardiac hypertrophy, and cancer. However, targeting these interactions specifically is nearly impossible in the absence of structural data that informs on the conformational differences between inactive, active, and G protein bound PLCβ states. The goal of this project is to understand mechanisms of PLCβ activation by Gβγ and/or the membrane through structural and functional studies. The Gβγ binding site on PLCβ has yet to be defined, and overall, Gβγ- dependent regulation of effector enzymes is a poorly understood process. This project would not only increase our overall understanding of PLCβ signaling at the membrane and in conjunction with Gβγ but would also provide insights into the regulation of other enzymes that act at the membrane interface. The completion of this study will synergistically increase our understanding of PLCβ activation mechanisms, providing insights that can be leveraged for the development of probes, and ultimately potential therapeutics, targeting G protein–PLCβ interactions.

项目概要 阿片类药物危机、心血管疾病和癌症是当今最紧迫的医疗保健危机 PLCβ 酶对于正常镇痛、心血管和健康至关重要。 此外，PLCβ 与其 G 蛋白之间的相互作用受到破坏。 激活剂已被证明可以改善阿片类药物诱导临床前模型的结果 然而，针对这些相互作用是有针对性的。 在缺乏表明构象差异的结构数据的情况下几乎不可能 在非活性、活性和 G 蛋白结合 PLCβ 状态之间。 通过结构和/或结构了解 Gβγ 和/或膜激活 PLCβ 的机制 PLCβ 上的 Gβγ 结合位点尚未确定，总体而言，Gβγ- 效应酶的依赖性调节是一个人们知之甚少的过程，这个项目不会。 只会增加我们对膜上 PLCβ 信号传导的整体理解 与 Gβγ 一起发挥作用，但也可以提供对作用于 Gβγ 的其他酶的调节的见解。 这项研究的完成将协同增加我们对膜界面的理解。 PLCβ 激活机制的研究，提供可用于开发的见解 探针，以及最终潜在的治疗方法，针对 G 蛋白-PLCβ 相互作用。