Purification and identification of two serine class proteinases from dog mast biochemically and immunologically similar to human proteinases tryptase and chymase

Arch Biochem Biophys. 1988 Apr;262(1):232-44. doi: 10.1016/0003-9861(88)90185-3.

Abstract

Serine class proteinases with trypsin-like and chymotrypsin-like specificity were purified from dog mastocytoma tissue. An antiserum was produced against the chymotrypsin-like proteinase. The antiserum reacted with mast cells in skin sections prepared from normal dogs consistent with the proteinase being a mast cell constituent. The antiserum also cross-reacted with the major chymotrypsin-like proteinase isolated from normal dog skin and partially cross-reacted with human skin chymase. No cross-reaction was detected with rat chymase. The trypsin-like proteinase from dog mastocytoma tissue was similar to tryptase isolated from human skin. It had a similar subunit structure, was not inhibited by many protein proteolytic enzyme inhibitors, bound to heparin, and reacted strongly with antiserum against human tryptase. Antiserum against human tryptase also reacted with mast cells in skin sections prepared from normal dog skin. No immunocytochemical labeling of rat skin mast cells was observed with anti-human tryptase. These studies establish the presence of a trypsin-like and chymotrypsin-like proteinase in dog skin mast cells and provide immunological evidence which suggests that both proteinases are more closely related to human than rat mast cell proteinases. These immunological and biochemical relationships are important when comparing the roles of these proteinases in different animals.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Chymases
  • Cross Reactions
  • Dogs
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunodiffusion
  • Kinetics
  • Mast Cells / enzymology*
  • Peptide Hydrolases / analysis*
  • Peptide Hydrolases / immunology
  • Serine Endopeptidases / analysis*
  • Serine Endopeptidases / immunology
  • Serine Endopeptidases / isolation & purification*
  • Skin / cytology
  • Skin / enzymology
  • Substrate Specificity

Substances

  • Peptide Hydrolases
  • tosylarginine methyl ester hydrolase
  • Serine Endopeptidases
  • Chymases