Isolation and characterization of the d1 domain of Pseudomonas aeruginosa nitrite reductase.
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.
Silvestrini
M C
MC
Dipartimento di Scienze Biochimiche, Università di Roma "La Sapienza," Italy.
Cutruzzolà
F
F
Schininà
M E
ME
Maras
B
B
Rolli
G
G
Brunori
M
M
D000595:Amino Acid Sequence; D002248:Carbon Monoxide / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D006418:Heme / Q000032:analysis; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009572:Nitrite Reductases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010782:Photolysis; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013053:Spectrophotometry; D013381:Subtilisins
D046911:Macromolecular Substances; D010446:Peptide Fragments; D006418:Heme; D002248:Carbon Monoxide; D009572:Nitrite Reductases; D010450:Endopeptidases; D013381:Subtilisins