title: Solid-state magic-angle spinning NMR of outer-membrane protein G from Escherichia coli.
abstract: Uniformly 13C-,15N-labelled outer-membrane protein G (OmpG) from Escherichia coli was expressed for structural studies by solid-state magic-angle spinning (MAS) NMR. Inclusion bodies of the recombinant, labelled protein were purified under denaturing conditions and refolded in detergent. OmpG was reconstituted into lipid bilayers and several milligrams of two-dimensional crystals were obtained. Solid-state MAS NMR spectra showed signals with an apparent line width of 80-120 Hz (including homonuclear scalar couplings). Signal patterns for several amino acids, including threonines, prolines and serines were resolved and identified in 2D proton-driven spin-diffusion (PDSD) spectra.
authors.affiliation: Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Hiller
authors.forename: Ludwig
authors.initials: L
authors.lastname: Krabben
authors.forename: Kutti R
authors.initials: KR
authors.lastname: Vinothkumar
authors.forename: Federica
authors.initials: F
authors.lastname: Castellani
authors.forename: Barth-Jan
authors.initials: BJ
authors.lastname: van Rossum
authors.forename: Werner
authors.initials: W
authors.lastname: Kühlbrandt
authors.forename: Hartmut
authors.initials: H
authors.lastname: Oschkinat
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003460:Crystallization; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D018272:Porins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011392:Proline / Q000737:chemistry; D017510:Protein Folding; D013912:Threonine / Q000737:chemistry
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C114347:OmpG protein, E coli; D018272:Porins; D013912:Threonine; D011392:Proline