Open AccessHomonuclear and heteronuclear NMR studies of oxidized Desulfovibrio vulgaris flavodoxin
Author(s) -
KNAUF Martin A.,
LÖHR Frank,
CURLEY G. Paul,
O'FARRELL Paul,
MAYHEW Stephen G.,
MÜLLER Franz,
RÜTERJANS Heinz
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb17746.x
Subject(s) - flavodoxin , heteronuclear molecule , desulfovibrio vulgaris , homonuclear molecule , chemistry , crystallography , protein secondary structure , desulfovibrio , nuclear overhauser effect , nuclear magnetic resonance spectroscopy , two dimensional nuclear magnetic resonance spectroscopy , stereochemistry , heteronuclear single quantum coherence spectroscopy , molecule , biochemistry , biology , ferredoxin , organic chemistry , bacteria , sulfate , genetics , enzyme
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. Sequencespecific assignment of all backbone and most of the side chain 1 H and 15 N 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 H N ‐H α spin‐spin coupling constants. In solution, flavodoxin consists of a five‐stranded parallel β‐sheet and four α‐helices. Residues 3–9, 32–36, 52–58, 87–96, and 123–128 are involved in the β‐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.