2h3o
Structure of MERFT, a membrane protein with two trans-membrane helices
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OverviewOverview
The structure of the membrane protein MerFt was determined in magnetically, aligned phospholipid bicelles by solid-state NMR spectroscopy. With two, trans-membrane helices and a 10-residue inter-helical loop, this truncated, construct of the mercury transport membrane protein MerF has sufficient, structural complexity to demonstrate the feasibility of determining the, structures of polytopic membrane proteins in their native phospholipid, bilayer environment under physiological conditions. PISEMA, SAMMY, and, other double-resonance experiments were applied to uniformly and, selectively (15)N-labeled samples to resolve and assign the backbone amide, resonances and to measure the associated (15)N chemical shift and, (1)H-(15)N heteronuclear dipolar coupling frequencies as orientation, constraints for structure calculations. (1)H/(13)C/(15)N triple-resonance, experiments were applied to selectively (13)C'- and (15)N-labeled samples, to complete the resonance assignments, especially for residues in the, nonhelical regions of the protein. A single resonance is observed for each, labeled site in one- and two-dimensional spectra. Therefore, each residue, has a unique conformation, and all protein molecules in the sample have, the same three-dimensional structure and are oriented identically in, planar phospholipid bilayers. Combined with the absence of significant, intensity near the isotropic resonance frequency, this demonstrates that, the entire protein, including the loop and terminal regions, has a, well-defined, stable structure in phospholipid bilayers.
About this StructureAbout this Structure
2H3O is a Single protein structure of sequence from Morganella morganii. Full crystallographic information is available from OCA.
ReferenceReference
Structure determination of a membrane protein with two trans-membrane helices in aligned phospholipid bicelles by solid-state NMR spectroscopy., De Angelis AA, Howell SC, Nevzorov AA, Opella SJ, J Am Chem Soc. 2006 Sep 20;128(37):12256-67. PMID:16967977
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