id="title" class="nomar"Membrane Protein Structures by Solution NMR
The MPSbyNMR center is a PSI membrane protein structure determination center, and consists of investigators from Harvard Medical School, Vanderbilt University, Frankfurt University, Los Alamos National Laboratory, and FB Reagents.
- Harvard Medical School
- Dana-Farber Cancer Institute, Harvard Medical School
- Vanderbilt University
- Frankfurt University
- Bioscience division of Los Alamos National Laboratory
- FB Reagents
A Protein Structure Initiative (PSI) Center that aims to develop an efficient solution NMR pipeline for solving membrane protein (MP) structures. To drive technology development and to test the proposed pipeline, we have selected 10 MP targets for which the structures are not known, including membrane-embedded transporters, enzymes, and receptors. These targets are polytopic helical MPs with 3-7 transmembrane helices and with sizes from 18 – 43 kDa. The Center will develop technologies that will have immediate and practical impact on structure determination. They include (1) cell-free expression platforms for production of MPs and for screening for NMR-feasible MP targets; (2) new detergents, bicelles and MP refolding methods; (3) non-uniform sampled high resolution 4D NOESYs; (4) new strategies for selective isotope labeling of methyl groups for acquiring long-range NOEs; (5) novel reagent for site-directed paramagnetic tagging and universal DNA-nanotube alignment media for RDC measurements; and (6) RDC-based molecular fragment replacement and structure calculation protocols. Although the phrase “high-throughput” does not yet apply to MP in any technological context, we aim to establish a NMR tool package within the proposed funding period that has the capacity for systematic production of MP structures.