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Identification of Porphyromonas gingivalis lipopolysaccharide-binding proteins in human saliva
Cited 34 time in
Web of Science
Cited 37 time in Scopus
- Authors
- Issue Date
- 2011-09
- Publisher
- PERGAMON-ELSEVIER SCIENCE LTD
- Citation
- MOLECULAR IMMUNOLOGY, Vol.48, No.15-16, pp.2207-2213
- Keywords
- Porphyromonas gingivalis ; Lipopolysaccharide ; Lipopolysaccharide-binding proteins ; Mass spectrometry ; Saliva
- Abstract
- Porphyromonas gingivalis causes periodontal diseases and its lipopolysaccharide (LPS) is considered as a major virulence factor responsible for pathogenesis. Since initial recognition of P. gingivalis LPS (Pg.LPS) in the oral cavity might be crucial for the host response, we identified Pg.LPS-binding proteins (Pg.LPS-BPs) using Pg.LPS-immobilized beads and a high-resolution mass spectrometry. LPS purified from P. gingivalis was conjugated onto N-hydroxysuccinimidyl-Sepharose (R) 4 Fast Flow beads. Notably, Pg.LPS-conjugated beads could stimulate Toll-like receptor 2 (TLR2) as determined by a TLR2-depdendent reporter expression system using CHO/CD14/TLR2. In addition, the Pg.LPS-conjugated beads induced the production of inflammatory mediators such as nitric oxide and interferon-gamma-inducible protein-10 in the macrophage cell-line, RAW 264.7. These results imply that Pg.LPS retained its immunological properties during the conjugation process. Then, the Pg.LPS-conjugated beads were mixed with a pool of saliva obtained from nine human subjects to capture Pg.LPS-BPs and molecular identities were determined by LTQ-Orbitrap hybrid fourier transform mass spectrometry. Pg.LPS-BPs captured at high frequencies included alpha-amylase, cystatin, prolactin-inducible protein, lysozyme C, immunoglobulin components, serum albumin, lipocalin-1, and submaxillary gland androgen-regulated protein 3B. These proteins are known to be involved in bacterial adhesion and colonization, anti-microbial functions or modulation of immune responses. (C) 2011 Elsevier Ltd. All rights reserved.
- ISSN
- 0161-5890
- Language
- English
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