title: Homonuclear and heteronuclear NMR studies of oxidized Desulfovibrio vulgaris flavodoxin. Sequential assignments and identification of secondary structure elements.
abstract: Recombinant Desulfovibrio vulgaris flavodoxin (molecular mass 16.3 kDa) was produced in Escherichia coli. The oxidized protein has been investigated with a combination of homonuclear and heteronuclear two-dimensional and heteronuclear three-dimensional NMR spectroscopy. Sequence-specific assignment of all backbone and most of the side chain 1H and 15N resonances has been obtained. The secondary structure has been inferred from the pattern of sequential, medium-, and long-range NOEs, together with information about slowly exchanging amide hydrogens and HN-H alpha spin-spin coupling constants. In solution, flavodoxin consists of a five-stranded parallel beta-sheet and four alpha-helices. Residues 3-9, 32-36, 52-58, 87-96, and 123-128 are involved in the beta-sheet whereas the a-helical regions comprise residues 13-28, 69-76, 104-114, and 134-148. Several proton resonances of the bound flavin mononucleotide cofactor have been assigned. NOE contacts between the prosthetic group and the apoprotein have been detected.
authors: Institute of Biophysical Chemistry, University Frankfurt, Germany.
authors: M A
authors: MA
authors: Knauf
authors: F
authors: F
authors: Löhr
authors: G P
authors: GP
authors: Curley
authors: P
authors: P
authors: O'Farrell
authors: S G
authors: SG
authors: Mayhew
authors: F
authors: F
authors: Müller
authors: H
authors: H
authors: Rüterjans
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D016969:Desulfovibrio vulgaris / Q000737:chemistry*; D005486:Flavin Mononucleotide / Q000378:metabolism; D005415:Flavins / Q000737:chemistry; D005418:Flavodoxin / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D010084:Oxidation-Reduction; D011392:Proline / Q000737:chemistry; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D012996:Solutions
chemical_list: D005415:Flavins; D005418:Flavodoxin; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D012996:Solutions; D005486:Flavin Mononucleotide; D011392:Proline