Homonuclear and heteronuclear NMR studies of oxidized Desulfovibrio vulgaris flavodoxin. Sequential assignments and identification of secondary structure elements.
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.
Knauf
M A
MA
Institute of Biophysical Chemistry, University Frankfurt, Germany.
Löhr
F
F
Curley
G P
GP
O'Farrell
P
P
Mayhew
S G
SG
Müller
F
F
Rüterjans
H
H
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
D005415:Flavins; D005418:Flavodoxin; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D012996:Solutions; D005486:Flavin Mononucleotide; D011392:Proline