Regulated expression, processing, and secretion of dog mast cell dipeptidyl peptidase I.
Dipeptidyl peptidase I (DPPI) is a cysteine protease found predominantly in myelomonocytic cells, cytotoxic T-cells, and mast cells. Recent studies identify an intracellular role for mast cell-DPPI (MC-DPPI) by activating prochymase and protryptase to their mature forms. To better define MC-DPPI and to explore the possibility of extracellular roles, we purified MC-DPPI from mastocytoma cells. We found the dog C2 mastocytoma cell line to be the richest source yet described for DPPI, purifying up to 200 microg of enzyme per g of cells. Dog MC-DPPI has an Mr of approximately 175,000 and consists of four subunits, each composed of a propeptide, light chain, and heavy chain. The heavy chain is N-glycosylated and is heterogeneously processed to three different forms. NH2-terminal sequences of the heavy chain and propeptide are identical to those predicted from a cDNA clone we sequenced from a mastocytoma cDNA library. The dog cDNA-derived sequence is 86% identical to that of human DPPI. Dog mastocytoma cells incubated with 12-O-tetradecanoylphorbol-13-acetate increase expression of MC-DPPI mRNA. MC-DPPI maintains its activity for dipeptide substrates at a neutral to alkaline pH. Cells stimulated with ionophore or substance P secrete MC-DPPI in parallel with the granule-associated mediators tryptase and histamine. Thus, dog mastocytoma cells secrete DPPI that is active at the pH of extracellular fluids, suggesting that MC-DPPI may act outside the cell.
Wolters
P J
PJ
Department of Medicine and the Cardiovascular Research Institute, University of California, San Francisco, California 94143-0911, USA.
Raymond
W W
WW
Blount
J L
JL
Caughey
G H
GH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D020789:Cathepsin C; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000235:genetics* / Q000378:metabolism; D004285:Dogs; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008407:Mast Cells / Q000201:enzymology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D020789:Cathepsin C