title: Assignment of oriented sample NMR resonances from a three transmembrane helix protein.
abstract: Oriented sample solid state NMR techniques have been routinely employed to determine the structures of membrane proteins with one or two transmembrane helices. For larger proteins the technique has been limited by spectral resolution and lack of assignment strategies. Here, a strategy for resonance assignment is devised and applied to a three transmembrane helix protein. Sequence specific assignments for all labeled transmembrane amino acid sites are obtained, which provide a set of orientational restraints and helix orientations in the bilayer. Our experiments expand the utility of solid state NMR in membrane protein structure characterization to three transmembrane helix proteins and represent a straightforward strategy for routinely characterizing multiple transmembrane helix protein structures.
authors: Institute for Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA; National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA.
authors: D T
authors: DT
authors: Murray
authors: National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA.
authors: I
authors: I
authors: Hung
authors: Institute for Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA; National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA; Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA. Electronic address: cross@magnet.fsu.edu.
authors: T A
authors: TA
authors: Cross
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D002478:Cells, Cultured; D003198:Computer Simulation; D007553:Isotope Labeling; D008051:Lipid Bilayers; D008081:Liposomes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary
chemical_list: D000596:Amino Acids; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D009587:Nitrogen Isotopes
keywords: Helix orientation; Lipid bilayer; Membrane protein; Membrane protein structure; Mycobacterium tuberculosis; Oriented sample solid state NMR; Resonance assignment