Isolation and characterization of the d1 domain of Pseudomonas aeruginosa nitrite reductase

J Inorg Biochem. 1996 May 1;62(2):77-87. doi: 10.1016/0162-0134(95)00090-9.

Abstract

Proteolitic digestion of nitrite reductase from Pseudomonas aeruginosa allows to obtain and purify a domain containing only the d1 heme and constituted by two noncovalently bound peptides. This d1 domain catayzes oxygen consumption, and binds carbon monoxide with a kinetic constant slightly higher than the parental dimeric holoenzyme. The capacity to oxidize the physiological substrate, cytochrome c551, is lost, even when the proteolytic c heme domain is added to this reaction mixture. This finding suggests that the two domains do not have a significant affinity for each other, and are kept together only by being part of the same polypeptide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carbon Monoxide / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Endopeptidases
  • Heme / analysis
  • Kinetics
  • Macromolecular Substances
  • Molecular Sequence Data
  • Molecular Weight
  • Nitrite Reductases / chemistry*
  • Nitrite Reductases / isolation & purification
  • Nitrite Reductases / metabolism
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification
  • Photolysis
  • Pseudomonas aeruginosa / enzymology*
  • Spectrophotometry
  • Subtilisins

Substances

  • Macromolecular Substances
  • Peptide Fragments
  • Heme
  • Carbon Monoxide
  • Nitrite Reductases
  • Endopeptidases
  • Subtilisins