Proteinases of rat mast cells. Peritoneal but not intestinal mucosal mast cells express mast cell proteinase 5 and carboxypeptidase A.
Six basic proteins of 26 to 38 kDa with isoelectric points (pI) > or = 8.5 were abundant in proteins separated by two-dimensional SDS-PAGE from adult rat peritoneal mast cells (MC). One was identified previously as rat mast cell proteinase (RMCP) 1, a chymase of 26 to 28 kDa, pI > 9.0. Microsequence analyses showed that two polypeptides of about 29 and 30 kDa had NH2 terminal amino acid sequences homologous to mouse MC proteinase 5 (MCP-5), whereas the amino terminals of the 33, 35, and 36 kDa proteins were homologous to MC carboxypeptidase A (MC-CPA). Rabbit Abs produced against synthetic peptides of the identified NH2 terminal sequences were used in immunoblot studies. At least three proteins reacted with Abs to MC-CPA, whereas Abs to MCP-5 detected three adjacent polypeptides, rather than just the two identified by using microsequence analysis. Removal of oligosaccharide side chains using peptide:N-glycosidase F reduced the heterogeneity of each set of three polypeptides (MCP-5 and MC-CPA) to a band of each protein of a lower M(r). The serine proteinase inhibitor [3H]diisopropylfluorophosphate ([3H]DFP) bound to a proteinase of 30 to 35 kDa, which is probably MC tryptase (pI < or = 6.0). Immunoblot analysis of proteins from intestinal mucosal mast cells showed RMCP-2, but not RMCP-1, MCP-5, or MC-CPA. This is the first report of MCP-5 in the rat and of clearly distinguishable glycosylated forms of MC CPA. These proteinases appear to be restricted in their distribution to selected MC populations, but little is known about their functions.
Befus
A D
AD
Department of Medicine, University of Alberta, Edmonton, Canada.
Chin
B
B
Pick
J
J
Evans
S
S
Osborn
S
S
Forstrom
J
J
D000581:Amidohydrolases; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis; D002268:Carboxypeptidases / Q000276:immunology / Q000378:metabolism*; D043422:Carboxypeptidases A / Q000276:immunology / Q000378:metabolism*; D053818:Chymases; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology* / Q000276:immunology; D008297:Male; D008407:Mast Cells / Q000201:enzymology* / Q000276:immunology; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010455:Peptides / Q000276:immunology; D010537:Peritoneum / Q000166:cytology / Q000201:enzymology* / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D053802:Tryptases
D000906:Antibodies; D010455:Peptides; C509190:Tpsb2 protein, mouse; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; C000593420:mast cell carboxypeptidase A, rat; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; C509222:Tpsab1 protein, mouse; C509223:Tpsab1 protein, rat; C509221:Tpsb2 protein, rat; D053802:Tryptases; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase