Structure and Dynamics of Membrane Proteins from NMR Orientational Constraints

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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.