title: Lysine-specific proteolytic activity responsible for forsythia detaching factor modification.
abstract: OBJECTIVE
  The objective of the present study was to clarify the lysine-specific proteolytic activity derived from periodontal pathogens responsible for Forsythia detaching factor (FDF) modification.
  
  
  METHODS
  The activity responsible for FDF modification in Tannerella forsythia and Porphyromonas gingivalis were evaluated by colorimetric assay using Ac-Arg-Ala-Lys-p-nitroaniline as a substrate. FDF modification in T. forsythia and P. gingivalis were evaluated by Western blotting using recombinant FDF (rFDF) as a substrate. Furthermore, the activity in GCF of 20 patients with periodontitis and 10 healthy subjects was also evaluated by colorimetric assay. Bacteria in subgingival plaque were detected using polymerase chain reaction.
  
  
  RESULTS
  The activity of both bacteria in colorimetric assay were 21.35 unit (P. gingivalis) and 3.61 unit (T. forsythia), respectively. Western blot analysis revealed that P. gingivalis was found to efficiently degrade rFDF and T. forsythia partially cleaved rFDF. The activity in GCF from patients with periodontitis (clinically healthy sites: CH, deep bleeding sites: DB and deep non-bleeding sites: DNB) was significantly higher than those from healthy subjects (healthy sites: H). Among the patients with periodontitis, the activity from CH was significantly lower than those from DB and DNB. T. forsythia was detected in 68.4% of DNB, in 78.4% of DB and in none of CH. P. gingivalis was detected in 63.2% of DNB, in 84.0% of DB and in 10.5% of CH. No bacterium was detected in healthy subjects.
  
  
  CONCLUSIONS
  The lysine-specific proteolytic activity responsible for FDF modification correlates with the presence of major periodontal pathogens.
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan. Electronic address: onisperi@dent.meikai.ac.jp.
authors.forename: Hidetomo
authors.initials: H
authors.lastname: Onishi
authors.affiliation: Department of Periodontics, School of Dentistry, Loma Linda University, CA 92350, USA.
authors.forename: Munehiko
authors.initials: M
authors.lastname: Ro
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Ishii
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Hideharu
authors.initials: H
authors.lastname: Otsuka
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yatabe
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Joichiro
authors.initials: J
authors.lastname: Hayashi
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Junichi
authors.initials: J
authors.lastname: Tatsumi
authors.affiliation: Division of Periodontology, Department of Oral Biology & Tissue Engineering, Meikai University School of Dentistry, 1-1 Keyakidai, Sakado-shi, Saitama 350-0283, Japan.
authors.forename: Kitetsu
authors.initials: K
authors.lastname: Shin
mesh_terms: D000328:Adult; D001426:Bacterial Proteins / Q000502:physiology*; D015153:Blotting, Western; D016022:Case-Control Studies; D002457:Cell Extracts / Q000737:chemistry*; D002478:Cells, Cultured; D003124:Colorimetry; D005260:Female; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D010518:Periodontitis / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000378:metabolism*; D059748:Proteolysis; D000070436:Tannerella forsythia / Q000378:metabolism*; D037521:Virulence Factors / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D002457:Cell Extracts; D037521:Virulence Factors; D008239:Lysine
keywords: Forsythia detaching factor; GCF; Lysine-specific proteolytic activity; Porphyromonas gingivalis; Tannerella forsythia