SspB Peptide Assay Reveals Saliva-Mediated Porphyromonas gingivalis Attachment

Tatsuro Ito^1,2*, Hidenobu Senpuku³, Takahiro Ichinosawa¹, Nana Ikematsu-Ito¹, Nao Kimura¹, Takehiko Shimizu^1,2

¹Department of Pediatric Dentistry, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
²Nihon University Research Institute of Oral Science, Chiba, Japan.
³Department of Bacteriology, National Institute of Infectious Diseases, Tokyo, Japan.

DOI: 10.4236/ojst.2015.511032                

Abstract

Background: Porphyromonas gingivalis is a major periodontal pathogen that binds efficiently to Streptococcus gordonii, which in turn binds to salivary agglutinin (gp340). The SspB of S. gordonii appears to mediate this association. We previously reported that the strepto-coccal SspB peptide analog, designated SspB (390-T400K-402), showed high binding activity with saliva. To understand the three-way interaction among S. gordonii, P. gingivalis and salivary gp340 as a unit, we established a peptide binding assay using SspB (390-T400K-402). Methods: The binding activity of the SspB (390-T400K-402) to P. gingivalis was detected by ELISA. Ninety-six well plates were coated with whole bacterial cell (P. gingivalis strains ATCC 33277, and W83; S. gordonii DL1) in Na₂CO₃ coating buffer. After blocking, bacterial cells were incubated with saliva or salivary agglutinin peptide (SRCRP2). Biotinylated SspB (390-T400K-402) was applied and incubated with 1:1000 streptoavidin-conjugated alkaline phosphatase. After development, A405 was recorded. Results: P. gingivalis 33277 showed the highest binding activity of the tested bacteria, whereas P. gingivalis W83, which was deficient in Mfa1 fimbriae, exhibited poor binding activity, as did S. gordonii. The binding of SspB (390-T400K-402) peptide in saliva- or SRCRP2-treated P. gingivalis was significantly higher than that in non-treated cells. Conclusion: The SspB (390-T400K-402) peptide binding assay revealed that initial attachment of P. gingivalis to the substrata of S. gordonii may require gp340-mediated SspB-Mfa1 interactions. The assay is available to assess the relationships among SspB, Mfa1 and salivary gp340 as a unit.

Keywords

SspB, Biofilm, Porphyromonas gingivalis, Saliva, gp340

Conflicts of Interest

The authors declare no conflicts of interest.

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