Identification of the functional domain in a coaggregation factor from Porphyromonas gingivalis
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概要
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Porphyromonas gingivalis has been implicated as a major pathogen in periodontal diseases. The coaggregation factor plays an important role in colonization of P. gingivalis in the subgingival area through aggregation with other extant oral microorganisms. We previously succeeded in gene cloning of the 40-kDa outer membrane protein (OMP) from P. gingivalis and identified the 40-kDa OMP as one of coaggregation factors. The mouse monoclonal antibody Pg-omp-A2, which inhibited the coaggregation activity of P. gingivalis, was successfully constructed. In order to develop a useful immunotherapy, it is essential to understand the functional domain expressing aggregation activity. The availability of recombinant 40-kDa OMP and the identification of the antigenic determinant recognized by Pg-omp-A2 will allow for the determination of the functional domain of the coaggregation factor. In this study, we used a phage-displayed epitope mapping system to examine the functional domain expressing aggregation activity. Peptide-displayed phage clones were isolated by biopanning using Pg-omp-A2 and determined DNA nucleotide sequence and then predicted the amino acid sequence of epitope. The amino acid sequence of epitope region was similar to the region of ^<96>IALDQTLGIP^<105> in 40-kDa OMP. Next, a series of 6 peptides (designated peptide A to F) in 40-kDa OMP was designed and chemically synthesized, and then we examined their immuno-reactivity by Pg-omp-A2. Pg-omp-A2 recognized the peptide C that contained the IALDQTLGIP amino acid sequence. Furthermore, the peptide C reduced the inhibitory activity of Pg-omp-A2 against the aggregation of P. gingivalis vesicles with Streptococcus gordonii. These findings suggest that the aggregation-associated domain of P. gingivalis 40-kDa coaggregation factor may be IALDQTLGIP.
- 日本大学の論文
著者
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Shibata Yasuko
Department Of Biochemistry And Research Institute Of Oral Science Nihon University School Of Dentist
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Shibata Yasuko
Department Of Biochemistry And Molecular Biology Nihon University School Of Dentistry At Matsudo:res
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Abiko Yoshimitsu
Department Of Biochemistry And Research Institute Of Oral Science Nihon University School Of Dentist
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Abiko Yoshimitsu
Department Of Biochemistry And Molecular Biochemistry Nihon University School Of Dentistry At Matsud
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Yamada Kazuaki
Department Of Biochemistry Nihon University School Of Dentistry At Matsudo
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Kiyama-Kishikawa Michiko
Department of Biochemistry and Research Institute of Oral Science, Nihon University School of Dentis
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Kiyama-kishikawa Michiko
Department Of Biochemistry And Research Institute Of Oral Science Nihon University School Of Dentist
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