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.

项目总结/摘要 我们的研究重点是了解单程膜蛋白的寡聚复合物。的 单程蛋白是最大的一类，占所有膜蛋白的20%以上。他们是极端的 生物学重要性：仅人类蛋白质组就含有2,000多种单通道膜蛋白， 是无数生理功能的核心这些单程膜的跨膜螺旋 蛋白质通常通过寡聚化和构象变化发挥关键作用。了解 这些现象对于理解生物过程中功能是至关重要的， 以及许多疾病的机制。 我的实验室用两个互补的项目研究单程膜蛋白的寡聚化 - 一个是关于阐明一个重要生物系统的结构-功能关系，第二个目标是 了解跨膜螺旋结合的一般原理。因为这些膜蛋白 系统很难用传统的结构化方法来研究，我们采用了一种方法， 实验方法与先进的计算建模。计算建模缓解了缺乏 实验结构，提供结构解释的现有实验数据和指导， 实验设计 我们的第一个项目的目标是研究细胞膜蛋白的结构组织， 分裂体，一种控制细菌细胞分裂的大型多蛋白质。虽然在以下方面取得了进展： 了解其组成部分及其作用，分割体的结构体系及其精确的 机制仍然知之甚少。解开这个组织对于理解 控制细菌分裂的机制。这些知识还可以支持新的 控制细菌生长的策略。 我们的第二个项目旨在使用蛋白质设计来了解跨膜螺旋寡聚化。的 主题是最常见的跨膜二聚化基序之一，GASright基序。GASright -这是 最为人所知的是血型糖蛋白A二聚体的折叠-其特征在于在其 界面，排列形成GxxxG和类似的图案。螺旋紧密接触，促进 形成弱的Cα-H O=C氢键网络。我们可以通过计算预测 GASright二聚体的结构及其相对稳定性。我们的下一个目标是通过调节 通过配体结合位点的设计在这些二聚体中进行二聚化和构象转换。这是 膜蛋白工程的一个几乎未探索的领域，与自然系统极其相关， 可能在合成生物学中有潜在的应用。