Understanding the molecular basis of transmembrane protein association

了解跨膜蛋白关联的分子基础

• 批准号: 10001573
• 负责人: Alessandro Senes
• 金额: $ 35.63万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10001573
• 关键词: Architecture; Area; Bacteria; Binding Sites; Biological; Biological Process; Biophysics; Cell division; Cell physiology; Complex; Computer Models; Computing Methodologies; Data; Defect; Development; Dimerization; Disease; Experimental Designs; GYPA gene; Goals; Growth; Human; Hydrogen Bonding; Integral Membrane Protein; Knowledge; Laboratory Study; Ligand Binding; Membrane Proteins; Methodology; Methods; Molecular; Molecular Conformation; Pattern; Physiological; Play; Protein Engineering; Proteins; Proteome; Research; Role; Structure; Structure-Activity Relationship; System; Testing; biological systems; design; dimer; human disease; membrane assembly; organizational structure; protein complex; synthetic biology; theories

PROJECT SUMMARY/ABSTRACT Our research focuses on understanding oligomeric complexes of single-pass membrane proteins. The single-pass proteins are the largest class, comprising over 20% of all membrane proteins. They are of extreme biological importance: the human proteome alone contains over 2,000 single-pass membrane proteins which are central to a myriad of physiological functions. The transmembrane helices of these single-pass membrane proteins often play a critical role through oligomerization and conformational change. Understanding the structural and biophysical basis these phenomena is critical to understanding function in biological processes, and the mechanisms of many diseases. My laboratory studies oligomerization of single-pass membrane proteins with two complementary projects – one related to elucidating structure-function relationship in an important biological system, the second aiming to understand the general principles of transmembrane helix association. Because these membrane protein systems are difficult to study with the traditional structural methods, we apply a methodology that integrates experimental methods with advanced computational modeling. The computational modeling mitigates the lack of experimental structure, providing structural interpretation of the available experimental data and guidance for experimental designs. The goal of our first project is to investigate the structural organization of membrane proteins of the divisome, the large multi-protein that governs cell division in bacteria. Although progress has been achieved in understanding its components and their roles, the structural architecture of the divisome and its precise mechanisms are still poorly understood. Unraveling this organization is crucial for understanding the mechanisms that govern bacterial division. This knowledge could also support the development of new strategies for controlling bacterial growth. Our second project seeks to understand transmembrane helix oligomerization using protein design. The subject is one of the most common transmembrane dimerization motifs, the GASright motif. GASright – which is best known as the fold of the glycophorin A dimer – is characterized by the presence of small amino at its interface, arranged to form GxxxG and similar patterns. The helices are in close contact, promoting the formation of networks of weak Cα–H∙∙∙O=C hydrogen bonds. We are able to predict computationally the structure of GASright dimers and their relative stability. Our next goal is to test our theories by modulating dimerization and conformational switching in these dimers through the design of ligand binding sites. This is an almost unexplored area of membrane protein engineering that is extremely relevant for natural systems and could potentially have applications in synthetic biology.

项目摘要/摘要 我们的研究重点是理解单通膜蛋白的寡聚复合物 单通蛋白是最大的类，成分超过所有膜蛋白。 生物学重要性：仅人类蛋白质组包含超过2000个单通膜蛋白 是无数生理功能的核心。 蛋白质通常通过寡聚和预约变化发挥关键作用。 结构和生物物理基础这些现象对于理解生物过程中的功能至关重要， 以及许多疾病的机制。 我的实验室研究通过两个互补项目将单通膜蛋白的低聚 - 与重要的生物系统中阐明结构 - 功能有关的一个，第二个目标 了解跨膜螺旋结合的一般原理。 系统是用传统结构方法研究的，我们应用了一种集成的方法 具有高级计算建模的实验方法。 实验结构，提供可用的体验数据的结构解释和指导 实验设计。 我们的项目的目的是调查膜蛋白的结构组织 Divisome，是政治Bactteria细胞分裂的大型多蛋白。 了解其组成部分及其角色，分裂的结构结构及其精确 机制仍然是理解的。 统治巴基的机制。 控制细菌生长的策略。 我们的第二个项目试图使用蛋白质设计来了解跨膜螺旋的寡聚化 受试者是最常见的跨膜二聚体基序之一，即气体基序。 最著名的是糖果蛋白a二聚体的折叠 - 特征是存在小氨基 界面，安排形成GXXXG和类似的模式。 弱cαH的网络的形成，我们要在计算上预测氢键。 气体二聚体的结构和相对稳定性。 这些二聚体中的二聚化和同意切换坐落在配体结合的设计。 膜蛋白工程几乎未开发的区域，与自然系统和 有可能在合成生物学中应用。