Analysis of the protein nano-environment of voltage-activated N-type Ca2+ channels Cav2.2 in the brain

脑内电压激活N型Ca2通道Cav2.2蛋白纳米环境分析

基本信息

批准号：
218389977
负责人：
Professor Dr. Bernd Fakler
金额：
--
依托单位：
Lehrstuhl für Physiologie II - Molecular Physiology
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2012
资助国家：
德国
起止时间：
2011-12-31 至 2015-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/218389977?language=en
关键词：
Analysis protein nano environment voltage

项目摘要

Voltage-gated N-type calcium channels (Cav2.2) trigger a variety of process in the CNS including function, formation and plasticity of synapses, activity-dependent gene expression and regulation of excitability and propagation of electrical signaling. Using comprehensive and quantitative proteomics on the molecular nano-environment of these channels we have identified the framework for their marked diversity in function: Cav2.2 channels are integrated into extended protein networks that may be assembled from a pool of ~160 proteins (at distinct abundance and stability of association) and reflect the cellular processes that can be fueled by Cav2.2 channels. The next funding period aims at the understanding (i) of the molecular structure and organization of the Cav2.2 channel networks, (ii) their local specificity and dynamics of assembly (do networks depend on the types of synapse/brain regions, on cellular boundaries such as neuronal activity and/or the presence of 'hub proteins'?), as well as on (iii) protein-protein interactions decisive for the operation of Cav2.2 nano-environments. For these goals we will apply a multidisciplinary approach comprising interactive application of an array of techniques including high-resolution quantitative mass spectrometry, biochemistry, standard molecular biology, immunocytochemistry and immunoelectron microscopy and electrophysiology.

电压门控 N 型钙通道 (Cav2.2) 触发 CNS 中的多种过程，包括突触的功能、形成和可塑性、活动依赖性基因表达以及兴奋性调节和电信号传播。通过对这些通道的分子纳米环境进行全面和定量的蛋白质组学分析，我们确定了其功能显着多样性的框架：Cav2.2 通道被整合到扩展的蛋白质网络中，该网络可以由约 160 种蛋白质（以不同的水平）组成的池组装而成。关联的丰度和稳定性）并反映由 Cav2.2 通道推动的细胞过程。下一个资助期旨在了解 (i) Cav2.2 通道网络的分子结构和组织，(ii) 它们的局部特异性和组装动力学（网络是否取决于突触/大脑区域的类型、细胞的类型）边界，例如神经元活动和/或“中心蛋白”的存在？），以及（iii）对 Cav2.2 纳米环境的运行起决定性作用的蛋白质-蛋白质相互作用。为了实现这些目标，我们将采用多学科方法，包括一系列技术的交互应用，包括高分辨率定量质谱、生物化学、标准分子生物学、免疫细胞化学、免疫电子显微镜和电生理学。