D. Claveau et D. Riendeau, Mutations of the C-terminal end of cathepsin K affect proenzyme secretion and intracellular maturation, BIOC BIOP R, 281(2), 2001, pp. 551-557
Citations number
30
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Transfection of the human cathepsin K cDNA into CHO cells results in the ex
pression of mature catalytically active 27-kDa protein and in cells secreti
ng the 39-kDa proenzyme form. Monensin, which neutralizes the pH of acidic
organelles, was found to inhibit intracellular processing of the proenzyme
and to stimulate its secretion into the culture medium. Brefeldin A caused
alterations in immunofluorescence staining consistent with interference of
lysosomal targeting and inhibited both intracellular processing and secreti
on of cathepsin K. Inhibition of glycosylation by tunicamycin also abolishe
d cathepsin K maturation. Furthermore, the processing of the proenzyme to t
he mature form was abolished by a single mutation of the terminal Met(329)
to Ala. The triple mutation of Ser(325) Pro(327), and Met(329) (all to Ala)
inhibited both maturation and secretion, using either transient or stable
expression systems, The results indicate that intracellular maturation and
secretion of cathepsin K can he affected differentially by various treatmen
ts and by mutations of the C-terminal end of the protein. These results are
consistent with the involvement of both the secreted proenzyme and the int
racellularly processed enzyme in cathepsin K-mediated processes. (C) 2001 A
cademic Press.