Analysis of the nifHDK operon and structure of the NifH protein from the unicellular, diazotrophic cyanobacterium, Cyanothece strain sp. ATCC 51142(1).
Cyanothece sp. ATCC 51142 is a unicellular, diazotrophic cyanobacterium that demonstrates diurnal rhythms for photosynthesis and N(2) fixation, with peaks of O(2) evolution and nitrogenase activity approximately 12 h out of phase. We cloned and sequenced the nifHDK operon, and determined that the amino acid sequences of all three proteins were highly conserved relative to those of other cyanobacteria and bacteria. However, the Fe-protein, encoded by the nifH gene, demonstrated two differences from the related protein in Azotobacter vinelandii, for which a 3-D structure has been determined. First, the Cyanothece Fe-protein contained a 37 amino acid extension at the N-terminus. This approximately 4 kDa addition to the protein appeared to fold as a separate domain, but remained a part of the active protein, as was verified by migration on acrylamide gels. In addition, the Cyanothece Fe-protein had amino acid differences at positions involved in formation of the Fe-protein dimer-dimer contacts in A. vinelandii nitrogenase. There were also changes in residues involved with interaction between the Fe-protein and the MoFe-protein when compared with A. vinelandii. Since the Cyanothece Fe-protein is quickly degraded after activity, it is suggested that the extension and the amino acid alterations were somehow involved in this degradative process.
Colón-López
M S
MS
Department of Biological Sciences, 1392 Lilly Hall of Life Sciences, Purdue University, West Lafayette, IN 47907-1392, USA.
Tang
H
H
Tucker
D L
DL
Sherman
L A
LA
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000737:chemistry / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D008983:Molybdoferredoxin / Q000737:chemistry / Q000235:genetics*; D009586:Nitrogen Fixation / Q000235:genetics; D009591:Nitrogenase / Q000737:chemistry*; D016366:Open Reading Frames; D009876:Operon; D010088:Oxidoreductases; D011487:Protein Conformation; D016415:Sequence Alignment
D008983:Molybdoferredoxin; D010088:Oxidoreductases; D009591:Nitrogenase; C029130:nitrogenase reductase