Resolving the mechanism of specific G-protein inhibitors by NMR spectroscopy

通过核磁共振波谱解析特定 G 蛋白抑制剂的机制

• 批准号: 419032556
• 负责人: Professor Dr. Clemens Glaubitz
• 金额: --
• 依托单位: Institut für Biophysikalische Chemie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/419032556?language=en
• 关键词: Resolving; mechanism; specific; protein; inhibitors

One possibility to disentangle the multiplicity of receptor-G protein interactions is to apply G-protein-specific inhibitors. So far, such inhibitors are rare but the depsipeptide FR900359 has been shown to act in a highly selective way onto the α-subunit of the G-Protein Gq. It also diminishes the activity of oncogenic Gqα mutants. Many details of the key step of the G-protein cycle, namely GDP/GTP exchange, and the mechanism of action of FR900359 remain to be resolved. Here, mainly solid-state NMR spectroscopy will be used to analyse Gqα in the membrane context by characterizing the mode of nucleotide binding, by analysing the intrinsic GTPase activity and by describing protein structure and dynamics in response to FR900359. The study will be expanded towards specific mutants, which are constitutively active and are considered as oncogenic drivers for various cancers.

一种可能解开受体- g蛋白相互作用的多样性是应用g蛋白特异性抑制剂。到目前为止，这样的抑制剂很少见，但已经证明，沉淀肽FR900359以高度选择性的方式作用于g蛋白Gq的α-亚基。它还能降低致癌Gqα突变体的活性。g蛋白周期的关键步骤，即GDP/GTP交换的许多细节，以及FR900359的作用机制仍有待解决。本文主要采用固态核磁共振波谱技术，通过表征核苷酸结合模式、分析GTPase活性以及描述响应FR900359的蛋白质结构和动力学来分析膜背景下的Gqα。这项研究将扩展到特定的突变体，这些突变体具有组成活性，被认为是各种癌症的致癌驱动因素。