title: Molecular engineering of a small trypsin inhibitor based on the binding loop of horsegram seed Bowman-Birk inhibitor.
abstract: BACKGROUND
  The Bowman-Birk inhibitors (BBIs) are currently investigated with renewed interest due to their therapeutic properties in cancer and other inflammatory disease treatment. The molecular mass of the BBI is a limitation, as sufficient amounts of the inhibitor do not reach the organs outside the gastrointestinal tract when administered orally.
  
  
  METHODS
  The anti-tryptic domain of HGI-III of horsegram (Dolichos biflorus) was cloned using the vector pET-20b (+) and expressed in E. coli BL21 (DE3) pLysS.
  
  
  RESULTS
  Kinetic analysis of this anti-tryptic peptide (recombinant trypsin inhibitory domain (rTID)) reveals that it is a potent inhibitor of trypsin and human tryptase. The K(i) (3.2 ± 0.17 × 10(-8) M) establishes a very high affinity to bovine trypsin. rTID inhibited human lung tryptase (IC(50) 3.78 ± 0.23 × 10(-7) M). The rTID is resistant to the digestive enzymes found in humans and animals.
  
  
  CONCLUSIONS
  These properties propagate further research on the use of rTID as a therapeutic for cancer and other related inflammatory diseases.
authors.affiliation: Department of Protein Chemistry and Technology, Central Food Technological Research Institute, Council of Scientific and Industrial Research, Mysore, India.
authors.forename: Deepa G
authors.initials: DG
authors.lastname: Muricken
authors.forename: Lalitha R
authors.initials: LR
authors.lastname: Gowda
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003001:Cloning, Molecular; D004220:Disulfides / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013329:Structure-Activity Relationship; D014357:Trypsin / Q000378:metabolism*; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000096:biosynthesis / Q000494:pharmacology*; D014361:Trypsin Inhibitors / Q000096:biosynthesis / Q000494:pharmacology*; D053802:Tryptases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D014361:Trypsin Inhibitors; D014357:Trypsin; D053802:Tryptases