title: Modification of forsythia detaching factor by gingival crevicular fluid in periodontitis.
abstract: OBJECTIVE
  Forsythia detaching factor (FDF) is a virulence factor of Tannerella forsythia detected as a mixture of the 60-kDa form of FDF and the 28-kDa C-terminal fragment (FDFc). The objective of the present study was to clarify the proteolytic activity of gingival crevicular fluid (GCF) from patients with periodontitis and healthy subjects using recombinant FDF (rFDF) as substrate.
  
  
  METHODS
  Eleven patients with periodontitis and 6 healthy subjects were recruited. Modification of rFDF and subsequent production of rFDFc by proteolytic activity of GCF was determined by Western blotting. Proteolytic activity of GCF was evaluated using an Ac-Arg-Ala-Lys-p-nitroaniline substrate. Correlation analysis between two different sets of variables was performed. Variables used in this analysis were proteolytic activity, clinical parameters, relative band density of rFDFc and those of rFDF.
  
  
  RESULTS
  Proteolytic activity in GCF was significantly higher in patients with periodontitis than in healthy subjects. Production of rFDFc was determined by treatment of rFDF with GCF from patients with periodontitis and with GCF from healthy subjects. Correlations between clinical parameters and proteolytic activity in GCF were significantly positive. On the other hand, correlations between relative band density of rFDFc or rFDF on Western blot and cleaving activity or clinical parameters were significantly negative.
  
  
  CONCLUSIONS
  The detected extend of GCF-activity generating rFDFc from rFDF and/or even further degrading rFDF correlates with severity of periodontitis.
authors.affiliation: Division of Periodontology, Department of Oral Biology and Tissue Engineering, School of Dentistry, Meikai University, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan. onisperi@dent.meikai.ac.jp
authors.forename: Hidetomo
authors.initials: H
authors.lastname: Onishi
authors.forename: Munehiko
authors.initials: M
authors.lastname: Ro
authors.forename: Joichiro
authors.initials: J
authors.lastname: Hayashi
authors.forename: Junichi
authors.initials: J
authors.lastname: Tatsumi
authors.forename: Namba
authors.initials: N
authors.lastname: Satomi
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yatabe
authors.forename: Shinichi
authors.initials: S
authors.lastname: Arakawa
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Izumi
authors.forename: Kitetsu
authors.initials: K
authors.lastname: Shin
mesh_terms: D000328:Adult; D000814:Aniline Compounds / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001439:Bacteroides / Q000201:enzymology*; D005260:Female; D005883:Gingival Crevicular Fluid / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010446:Peptide Fragments / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D017622:Periodontal Attachment Loss / Q000378:metabolism; D010514:Periodontal Pocket / Q000378:metabolism; D010518:Periodontitis / Q000382:microbiology*; D059748:Proteolysis; D011994:Recombinant Proteins; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D000814:Aniline Compounds; D001426:Bacterial Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D037521:Virulence Factors; C008625:nitroaniline; D010447:Peptide Hydrolases