Sheep mast cell proteinase-1, a serine proteinase with both tryptase- and chymase-like properties, is inhibited by plasma proteinase inhibitors and is mitogenic for bovine pulmonary artery fibroblasts.
Sheep mast cell proteinase-1 (sMCP-1), a serine proteinase with dual chymase/tryptase activity, is expressed in gastrointestinal mast cells, and released systemically and on to the mucosal surface during gastrointestinal nematode infection. The potential for native plasma proteinase inhibitors to control sMCP-1 activity was investigated. Sheep alpha1-proteinase inhibitor (alpha1PI) inhibited sMCP-1 slowly, with second-order association rate constant (kass) 1. 1x10(3) M-1.s-1, whereas sheep contrapsin inhibited trypsin (kass 2.2x10(6) M-1.s-1) but not sMCP-1. Western-blot analysis and gel filtration showed that when added to serum or plasma, sMCP-1 was partitioned between alpha1PI and alpha2-macroglobulin. The possibility that significant cleavage of plasma proteins could occur before sMCP-1 was inhibited was investigated using gel filtration and SDS/PAGE after adding sMCP-1 to plasma. Cleavage of ovine fibrinogen occurred in the presence of excess alpha1PI and alpha2-macroglobulin, the alpha-chain being cleaved C-terminally and the beta-chain at the putative Lys-27. In addition, sMCP-1 was found to be mitogenic for bovine pulmonary artery fibroblasts, but was not mitogenic in the presence of soya-bean trypsin inhibitor. In terms of fibrinogen cleavage and fibroblast stimulation, sMCP-1 shows functional similarities to mast cell tryptase.
Pemberton
A D
AD
Department of Veterinary Clinical Studies, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Veterinary Field Station, Easter Bush, Roslin, Midlothian EH25 9RG, Scotland, U.K.
Belham
C M
CM
Huntley
J F
JF
Plevin
R
R
Miller
H R
HR
D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D053818:Chymases; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D008407:Mast Cells / Q000201:enzymology*; D008934:Mitogens / Q000494:pharmacology*; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000097:blood / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000166:cytology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000494:pharmacology*; D015843:Serpins; D012756:Sheep / Q000378:metabolism*; D013045:Species Specificity; D014361:Trypsin Inhibitors / Q000097:blood / Q000494:pharmacology; D053802:Tryptases; D000515:alpha 1-Antitrypsin / Q000494:pharmacology
D001798:Blood Proteins; D008934:Mitogens; D011480:Protease Inhibitors; D015843:Serpins; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; C033691:contrapsin; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases