Structure and Dynamics of Membrane Proteins from NMR Orientational Constraints

描述(申请人提供)：据预测，近30%的人类蛋白质组由膜蛋白组成。尽管它们具有重要的功能和出现的频率，但目前只有100种独特的膜蛋白被结构鉴定并被纳入PDB数据库。它们的包裹体水平低是因为它们既不能进行X射线结晶学结晶，也不能通过核磁共振光谱产生成功确定结构所需的常规NOE数据。残留偶极耦合(RDC)是从溶液和固体核磁共振波谱获得的另一种数据来源，可以从水或膜蛋白中获得相对丰富的数据。RDC是蛋白质结构和内部动力学的准确报告者。然而，主要基于RDC数据的结构确定协议是具有挑战性的，因为它们需要新的分析方法，这些方法的操作方式与使用NOE数据的方法完全不同。为了克服这一点，我们建议开发一种方法，使用RDC数据作为主要数据来源来确定蛋白质的结构。我们的主要目标是提供一个程序(名为REDCRAFT)，旨在满足主要基于RDC数据快速和准确确定膜蛋白结构和运动特征的需求。在REDCRAFT的测试和评估过程中，我们将获得RDC数据，并确定两个医学相关蛋白的结构：BACE1和PINK1。这些蛋白质分别与阿尔茨海默病(AD)和帕金森病(PD)有关。

DESCRIPTION (provided by applicant): Nearly 30% of the human proteome is predicted to consist of membrane proteins. Despite their functional importance and frequency of occurrence, only a ~100 unique membrane proteins have been structurally characterized and included in the PDB database. Their low level of inclusion is because they neither crystallize for X-ray crystallography nor produce the conventional NOE data through NMR spectroscopy that has been required for successful structure determination. Residual dipolar couplings (RDC), an alternative source of data obtained from solution and solid state NMR spectroscopy, can be acquired in relative abundance from aqueous or membrane proteins. RDCs are accurate reporters of protein structure and internal dynamics. However, structure determination protocols based primarily on RDC data are challenging, since they require new analysis methods that operate in fundamentally different ways than those that use NOE data. To overcome this, we propose the development of a methodology leading to structure determination of proteins using RDC data as the primary source of data. Our primary goal is delivery of a program (named REDCRAFT) designed to address the need for rapid and accurate determination of membrane protein structures and characterization of motion primarily based on RDC data. During the course of testing and evaluation of REDCRAFT we will acquired RDC data and determine the structure of two medically relevant proteins: BACE1 and PINK1. These proteins are involved in the Alzheimer's disease (AD) and Parkinson's disease (PD) respectively.