Purification and characterization of a novel tripeptidyl aminopeptidase from Streptomyces lividans 66.
An extracellular tripeptidyl aminopeptidase has been purified from Streptomyces lividans 66 cell-free cultures. The enzyme is a major component of the secreted proteolytic activity. The protease removes only the N-terminal tripeptide from recombinant human GM-CSF and IL-3 but does not cleave recombinant human IL-6. The enzyme cleaves the synthetic tripeptide substrates APA-pNA and APM-pNA but does not cleave substrates with blocked amino terminals. Smaller substrates are not cleaved. The enzyme appears to be a serine protease of 55 kDa molecular weight. The pH optimum is between 7.5 and 8.5 but varies slightly with the substrate. The N-terminal sequence and amino acid composition have been determined.
Krieger
T J
TJ
Cangene Corporation, Mississauga, Ontario, Canada.
Bartfeld
D
D
Jenish
D L
DL
Hadary
D
D
D000595:Amino Acid Sequence; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D007377:Interleukin-3 / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
D007375:Interleukin-1; D007377:Interleukin-3; D009842:Oligopeptides; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D010450:Endopeptidases; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases