Identification of an interleukin-1 beta converting enzyme-like activity that increases upon treatment of P19 cells with retinoic acid as the proteasome.
We examined changes in proteinase activities in P19 embryonal carcinoma cells during retinoic acid-induced differentiation. The interleukin-1 beta converting enzyme (ICE)-like Ac-YVAD-MCA hydrolytic activity was increased about 6-fold by treatment with retinoic acid. This activity was inhibited by N-ethylmaleimide and Ac-YVAD-H but not by E-64, EDTA, PMSF, or amastatin. The ICE-like activity in P19 cells eluted as a single peak just after the void volume on gel filtration. No ICE-like activity was observed at a molecular mass of 30-50 kDa. Enzymatic purification, Western blot analysis, and an immunoabsorption study demonstrated that the ICE-like activity in P19 cells is caused by the proteasome, and is stimulated during retinoic acid-induced differentiation. The proteasome purified from mouse liver also cleaved Ac-YVAD-MCA. These results strongly suggest that the proteasome is a major ICE-like proteinase in P19 cells and may be involved in the neural differentiation and the apoptotic pathway.
Kobayashi
T
T
Department of Neuromuscular Research, Department of Development and Differentiation, National Institute of Neuroscience, NCNP Ogawahigashi-cho, Tokyo.
Shinozaki
A
A
Momoi
T
T
Arahata
K
K
Tsukahara
T
T
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D018236:Carcinoma, Embryonal / Q000201:enzymology* / Q000473:pathology; D020170:Caspase 1; D002454:Cell Differentiation / Q000187:drug effects; D003374:Coumarins / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D008099:Liver / Q000201:enzymology; D051379:Mice; D009097:Multienzyme Complexes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry; D046988:Proteasome Endopeptidase Complex; D013379:Substrate Specificity; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology
D003374:Coumarins; D009097:Multienzyme Complexes; D009842:Oligopeptides; C096022:acetyl-tyrosyl-valyl-alanyl-aspartyl-7-amino-4-methylcoumarinamide; D014212:Tretinoin; D004492:Edetic Acid; D003546:Cysteine Endopeptidases; D020170:Caspase 1; D046988:Proteasome Endopeptidase Complex; D005033:Ethylmaleimide