The influence of differential processing of procathepsin H on its aminopeptidase activity, secretion and subcellular localization in human cell lines.
Cathepsin H is a unique member of the cysteine cathepsins that acts primarily as an aminopeptidase. Like other cysteine cathepsins, it is synthesized as an inactive precursor and activated by proteolytic removal of its propeptide. Here we demonstrate that, in human cells, the processing of the propeptide is an autocatalytic, multistep process proceeding from an inactive 41kDa pro-form, through a 30kDa intermediate form, to the 28kDa mature form. Tyr87P and Gly90P were identified as the two major endopeptidase cleavage sites, converting the 30kDa form into the mature 28kDa form. The level of processing differs significantly in different human cell lines. In monocyte-derived macrophages U937 and prostate cancer cells PC-3, the 28kDa form is predominant, whereas in osteoblasts HOS the processing from the 30kDa form to the 28kDa form is significantly lower. The aminopeptidase activity of the enzyme and its subcellular localization are independent of the product, however the 30kDa form was not secreted in HOS cells. The activity of the resulting cathepsin H in U937 cells was significantly lower than that in HOS cells, presumably due to the high levels of endogenous cysteine protease inhibitor cystatin F present specifically in this cell line. These results provide an insight into the dependence of human cathepsin H processing and regulation on cell type.
Department of Pharmaceutical Biology, University of Ljubljana, Ljubljana, Slovenia.
Matija
M
Rojnik
Zala R
ZR
Jevnikar
Bojan
B
Doljak
Samo
S
Turk
Nace
N
Zidar
Janko
J
Kos
D020816:Amino Acid Motifs; D056655:Cathepsin H / Q000737:chemistry / Q000378:metabolism*; D002403:Cathepsins / Q000737:chemistry / Q000378:metabolism*; D015891:Cystatins / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D004792:Enzyme Precursors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000201:enzymology / Q000378:metabolism*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010006:Osteoblasts / Q000201:enzymology / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D020298:U937 Cells
D015891:Cystatins; D004792:Enzyme Precursors; C550521:cystatin F, mouse; D002403:Cathepsins; C061923:preprocathepsin H; C534126:CTSH protein, human; D056655:Cathepsin H