Changes of lysosomal proteinase activities and their expression in rat cultured keratinocytes during differentiation.
The cathepsins B, H and L, lysosomal cysteine proteinases, play a major role in intracellular protein degradation. These proteinase activities and expressions were examined in a Ca2+ regulated epidermal culture system which consists of two morphological cell types: undifferentiated cells grown in low Ca2+ (0.1 mM concentration) and differentiated cells grown in high Ca2+ (1.8 mM concentration), respectively. Cathepsin B and L activities of the differentiated cells showed a several-fold increase compared to that of the undifferentiated cells. In addition, by using CM-cellulose column chromatography, cathepsin B and L were separated and the level of cathepsin L activity increased significantly. Cathepsin B, L and H were also detected by using an immunoblotting procedure in which their bands were expressed after differentiation was induced by the increasing calcium concentration. Cathepsin L activity and immunostaining intensity reached a maximum at 1 or 2 days of differentiation. In contrast, cystatin alpha (an endogenous inhibitor of cysteine-dependent cathepsins) appeared in the final stage of differentiation. These results indicate that the expression of epidermal cathepsins and their endogenous inhibitor are involved in part of the program of cell differentiation and the terminal differentiation process in cultured rat keratinocytes.
Tanabe
H
H
Department of Plastic and Reconstructive Surgery, St. Marianna University School of Medicine, Kawasaki, Japan.
Kumagai
N
N
Tsukahara
T
T
Ishiura
S
S
Kominami
E
E
Nishina
H
H
Sugita
H
H
D000818:Animals; D002401:Cathepsin B / Q000378:metabolism; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003374:Coumarins / Q000378:metabolism; D015891:Cystatins / Q000378:metabolism; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism*; D015151:Immunoblotting; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats
D003374:Coumarins; C070572:Csta protein, rat; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D011480:Protease Inhibitors; C021159:benzyloxycarbonyl-phenylalanylarginine-4-methylcoumaryl-7-amide; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534280:Ctsl protein, rat