Soybean Bowman-Birk protease inhibitor is a highly effective inhibitor of human mast cell chymase

Arch Biochem Biophys. 1997 Aug 1;344(1):133-8. doi: 10.1006/abbi.1997.0182.

Abstract

Soybean Bowman-Birk protease inhibitor (BBI) is an inhibitor of serine proteases with two functional inhibitory domains of different specificities: one is specific for chymotrypsin-like proteases, the other for trypsin-like proteases. Chymase and tryptase are serine proteases which are stored in mast cell granules and released upon degranulation. This work investigated the inhibition of human chymase and tryptase by BBI. Active-site titration of human skin chymase by BBI demonstrated that BBI was a highly effective inhibitor of human chymase. Virtually stoichiometric inhibition of chymase by BBI was observed at 10 nM chymase. Kinetic studies of the inhibition reaction yielded an association rate constant of 4.0 x 10(5) M(-1) s(-1) and a dissociation rate constant of 1.7 x 10(-5) s(-1). From these two constants we estimate a K(i) of 50 pM. Chymase/BBI complexes did not dissociate in SDS-PAGE analyses under nonreducing conditions, consistent with the formation of a very tight complex with little tendency to dissociate. In contrast to chymase, human tryptase was not inhibited by BBI. These studies demonstrate that BBI is a good inhibitor of human chymase, exhibiting reaction properties better than physiological inhibitors described to date.

Publication types

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

MeSH terms

  • Binding Sites
  • Blotting, Western
  • Chymases
  • Chymotrypsin / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Kinetics
  • Mast Cells / drug effects
  • Mast Cells / enzymology*
  • Protein Binding
  • Recombinant Proteins / metabolism
  • Serine Endopeptidases / metabolism*
  • Serine Proteinase Inhibitors / pharmacology*
  • Skin / enzymology
  • Trypsin Inhibitor, Bowman-Birk Soybean / pharmacology*
  • Tryptases

Substances

  • Recombinant Proteins
  • Serine Proteinase Inhibitors
  • Trypsin Inhibitor, Bowman-Birk Soybean
  • Serine Endopeptidases
  • chymase 2
  • Chymotrypsin
  • Chymases
  • Tryptases