title: Pancreatic secretory trypsin inhibitor acts as an effective inhibitor of cysteine proteinases gingipains from Porphyromonas gingivalis.
abstract: OBJECTIVE
  Porphyromonas gingivalis has been implicated as the major pathogen of periodontitis in adults. This organism produces an array of virulence factors, of which cysteine proteinases, referred to as gingipains K and R, are believed to play a crucial role in pathogenicity. The aim of this study was to investigate the susceptibility of gingipains K and R to inhibition by a pancreatic secretory trypsin inhibitor.
  
  
  METHODS
  Enzyme activities were measured spectrophotometrically using chromogenic turnover substrates. To estimate the value of the association constant (Ka), constant amounts of enzyme were reacted with increasing amounts of inhibitor to reach equilibrium. The Ka was calculated by fitting the experimental data to the given equation.
  
  
  RESULTS
  In this study it was shown that gingipains are susceptible to pancreatic Kazal-type trypsin inhibitors (pancreatic secretory trypsin inhibitors). Bovine pancreatic secretory trypsin inhibitor, having an Arg residue at the P1 position of the reactive site, specifically inhibited the activity of the Arg-specific cysteine proteinase gingipain R, whereas porcine inhibitor, possessing a Lys residue at the P1 position, exhibited activity only against the Lys-specific cysteine proteinase gingipain K. The Ka values for the inhibitor-proteinase interaction were 1.6 x 10(6) m(-1) and 2.0 x 10(4) m(-1) for gingipain R and gingipain K, respectively.
  
  
  CONCLUSIONS
  This finding is the first demonstration of the inhibitory potency of the Kazal-type specific trypsin inhibitors against cysteine proteinases. These discoveries open new possibilities for the use of naturally occurring inhibitors, displaying activity across enzyme families, as a model in designing new molecules of therapeutic significance.
authors.affiliation: Department of Food Hygiene and Consumer Protection, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, Wroclaw, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Bania
authors.forename: A
authors.initials: A
authors.lastname: Kubiak
authors.forename: K
authors.initials: K
authors.lastname: Wojtachnio
authors.forename: A
authors.initials: A
authors.lastname: Polanowski
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002645:Chickens; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000080867:Gingipain Cysteine Endopeptidases; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D013552:Swine; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000302:isolation & purification / Q000378:metabolism*; D037521:Virulence Factors
chemical_list: D018829:Adhesins, Bacterial; D015853:Cysteine Proteinase Inhibitors; D000080867:Gingipain Cysteine Endopeptidases; D037521:Virulence Factors; D014359:Trypsin Inhibitor, Kazal Pancreatic; D003546:Cysteine Endopeptidases