Nitrate reductase of Escherichia coli: completion of the nucleotide sequence of the nar operon and reassessment of the role of the alpha and beta subunits in iron binding and electron transfer.
The nucleotide sequence of the narGHJI operon that encodes the nitrate reductase of Escherichia coli was completed. It encodes four polypeptides NarG, NarH, NarJ and NarI of molecular weight 138.7, 57.7, 26.5 and 25.5 kDa, respectively. The analysis of deduced amino acid sequence failed to reveal any structure capable of binding iron within the NarG polypeptide. In contrast, cysteine arrangements typical of iron-sulfur centers were found in the NarH polypeptide. This suggested that the latter is an electron transfer unit of the nitrate reductase complex. Such a view is opposite to the current description of the nitrate reductase. The findings allowed us to propose a model for the electron transfer steps that occur during nitrate reduction. The NarG polypeptide was found to display a high degree of homology with numerous E. coli molybdoproteins. Moreover, the same genetic and functional organizations as well as the presence of highly conserved stretches of amino acids were noted between both NarG/NarH and DmsA/DmsB (encoding the dimethyl sulfoxide reductase) pairs.
Blasco
F
F
Groupe de biomembranes, L.C.B. UER de Luminy, Marseille, France.
Iobbi
C
C
Giordano
G
G
Chippaux
M
M
Bonnefoy
V
V
D000595:Amino Acid Sequence; D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D009565:Nitrate Reductases / Q000235:genetics*; D009876:Operon; D010084:Oxidation-Reduction; D011485:Protein Binding; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D012984:Software
D003062:Codon; D004269:DNA, Bacterial; D007501:Iron; D009565:Nitrate Reductases