Isolation and characterization of an enkephalin-hydrolyzing dipeptidyl-aminopeptidase from calf-brain striatum.
Cytosolic dipeptidyl-aminopeptidase with a high affinity for Leu-enkephalin (Km = 5-7 microM) was partially purified from the 25,000 g supernatant of calf-brain striatum. The procedure included pH 4.5 denaturation, DEAE-cellulose chromatography and Blue Sepharose CL-6B chromatography and resulted in preparations that are free from other enkephalin-hydrolyzing enzymes. This enzyme, which is called enkephalinase B, has a positively charged group in its active site and presumably also a Zn atom since the loss in activity induced by EDTA treatment can be restored without loss of substrate affinity by low concentrations of ZnSO4.
D000818:Animals; D001051:Apoenzymes / Q000032:analysis; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D003342:Corpus Striatum / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000032:analysis* / Q000302:isolation & purification; D010450:Endopeptidases / Q000032:analysis; D004745:Enkephalins / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008670:Metals / Q000032:analysis; D013379:Substrate Specificity
D001051:Apoenzymes; D004745:Enkephalins; D008670:Metals; D010450:Endopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C100152:dipeptidyl peptidase III