Inhibition of Porphyromonas gingivalis hemagglutinating activity by synthetic peptides derived from phage display selection using MAb against the recombinant outer membrane protein

Wei Jen Chang, Michiko Kishikawa-Kiyama, Yasuko Shibata, Sheng Yang Lee, Yoshimitsu Abiko

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Porphyromonas gingivalis has been implicated as an pathogen in the development of periodontitis, and hemagglutinins have been identified as an important adhesion onto the gingival tissue cells, and to attach and lyse erythrocytes to uptake Fe ion as an essential nutriant. The 40-kDa outer membrane protein (OMP) has been moleculary cloned from P. gingivalis 381. Since the antibody against recombinant (r) 40-kDa OMP inhibited the hemagglutinating activity, and the polymeric form of r40-kDa OMP itself expressed hemagglutinating activity, the 40-kDa OMP is thought to be one of the hemagglutinins. Moreover, we established MAbs against r40-kDa OMP which were capable of inhibiting hemagglutinating activity of P. gingivalis vesicles. In the present study, a phage-displayed epitope mapping system was used to identify the functional domain expressing hemagglutinating activity by biopanning using the neutralizing mAb, Pg-ompA1. The minimal epitope requirements of the MAb and the predicted amino acid sequences were identified in the region of 96IALDQTLGIP105 in 40-kDa OMP. Synthetic peptide, 87WPRVGQLFIALDQTLGIPTFSVCRME116, mapped the relevant molecule within a short stretch and is corresponding to residues of 40-kDa OMP. Chemically synthesized peptide was used to determine its inhibitory activity against hemagglutinating activity. The synthetic peptide significantly abolished hemagglutinating activity in a dose-dependent manner. These findings suggest that the synthetic peptide is an effective antagonist of erythrocyte binding, and this peptide may be a potent inhibitor of hemagglutination of P. gingivalis cells. The use of synthetic peptide neutralizing hemagglutinating activity of P. gingivalis represents a possible new therapeutic approach to P. gingivalis infected periodontitis.

Original languageEnglish
Pages (from-to)352-356
Number of pages5
JournalHybridoma and Hybridomics
Volume23
Issue number6
DOIs
Publication statusPublished - Dec 2004

Fingerprint

Porphyromonas gingivalis
Bacteriophages
Membrane Proteins
Peptides
Periodontitis
Hemagglutinins
Erythrocytes
Epitope Mapping
Hemagglutination
Epitopes
Amino Acid Sequence
Ions
Antibodies

ASJC Scopus subject areas

  • Genetics
  • Immunology

Cite this

Inhibition of Porphyromonas gingivalis hemagglutinating activity by synthetic peptides derived from phage display selection using MAb against the recombinant outer membrane protein. / Chang, Wei Jen; Kishikawa-Kiyama, Michiko; Shibata, Yasuko; Lee, Sheng Yang; Abiko, Yoshimitsu.

In: Hybridoma and Hybridomics, Vol. 23, No. 6, 12.2004, p. 352-356.

Research output: Contribution to journalArticle

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