Identification of an elastase-like activity, that decreased during the differentiation of MEL cells, as a prolyl endopeptidase.
1. A Suc-APA-MCA hydrolytic activity was significantly decreased in murine erythroleukemia cells during DMSO-induced differentiation, but not in DMSO-resistant cells. 2. The Suc-APA-MCA hydrolytic enzyme was purified by ion exchange, adsorption, gel filtration and affinity chromotographies. The results of the chromatographies showed that only one enzyme hydrolyzed Suc-APA-MCA in MEL cells. 3. This enzyme is more sensitive to hydrolysis by Suc-GPLGP-MCA than Suc-APA-MCA at slightly acidic pH, and its activity is stimulated by 2-mercaptoethanol. 4. A cysteine proteinase inhibitor did not affect the activity, but a specific inhibitor of prolyl endopeptidase, Z-thioprothiazolidine, completely inhibited it. These results suggest that the Suc-APA-MCA hydrolytic enzyme is identical to a prolyl endopeptidase.
Tsukahara
T
T
National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan.
Ishiura
S
S
Sugita
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004351:Drug Resistance; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D004915:Leukemia, Erythroblastic, Acute; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000302:isolation & purification / Q000378:metabolism*; D000085822:Prolyl Oligopeptidases; D011480:Protease Inhibitors / Q000494:pharmacology; D012697:Serine Endopeptidases; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
D011480:Protease Inhibitors; D008623:Mercaptoethanol; D010450:Endopeptidases; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases; D010196:Pancreatic Elastase; D003546:Cysteine Endopeptidases; D004121:Dimethyl Sulfoxide