Differential profiles of salivary proteins with affinity to Streptococcus mutans lipoteichoic acid in caries-free and caries-positive human subjects.
Streptococcus mutans is a representative oral pathogen that causes dental caries and pulpal inflammation. Its lipoteichoic acid (Sm.LTA) is known to be an important cell-wall virulence factor involved in bacterial adhesion and induction of inflammation. Since Sm.LTA-binding proteins (Sm.LTA-BPs) might play an important role in pathogenesis and host immunity, we identified the Sm.LTA-BPs in the saliva of caries-free and caries-positive human subjects using Sm.LTA-conjugated beads and LTQ-Orbitrap hybrid Fourier transform mass spectrometry. Sm.LTA was conjugated to N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads (Sm.LTA-beads). Sm.LTA retained its biological properties during conjugation, as determined by the expression of nitric oxide and interferon-γ-inducible protein 10 in a murine macrophage cell line and activation of Toll-like receptor 2 (TLR2) in CHO/CD14/TLR2 cells. Sm.LTA-BPs were isolated from pooled saliva prepared from 10 caries-free or caries-positive human subjects each, electrophoresed to see their differential expression in each group, and further identified by high-resolution mass spectrometry. A total of 8 and 12 Sm.LTA-BPs were identified with statistical significance in the pooled saliva from the caries-free and caries-positive human subjects, respectively. Unique Sm.LTA-BPs found in caries-free saliva included histone H4, profilin-1 and neutrophil defensin-1, and those in caries-positive saliva included cystatin-C, cystatin-SN, cystatin-S, cystatin-D, lysozyme C, calmodulin-like protein 3 and β-actin. The Sm.LTA-BPs found in both groups were hemoglobin subunits α and β, prolactin-inducible protein, protein S100-A9, and SPLUNC2. Collectively, we identified Sm.LTA-BPs in the saliva of caries-free and caries-positive subjects, which exhibit differential protein profiles.
Department of Oral Microbiology and Immunology, DRI, and BK21 Plus Program, School of Dentistry, Seoul National University, Seoul, Korea.
S W
SW
Hong
D-G
DG
Seo
J E
JE
Baik
K
K
Cho
C-H
CH
Yun
S H
SH
Han
D000199:Actins / Q000032:analysis; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology; D016466:CHO Cells; D002147:Calmodulin / Q000032:analysis; D002460:Cell Line; D054357:Chemokine CXCL10 / Q000187:drug effects; D003412:Cricetulus; D055316:Cystatin C / Q000032:analysis; D015891:Cystatins / Q000032:analysis; D023082:Defensins / Q000032:analysis; D003731:Dental Caries / Q000378:metabolism* / Q000382:microbiology; D006657:Histones / Q000032:analysis; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000187:drug effects; D008070:Lipopolysaccharides / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects; D051379:Mice; D009113:Muramidase / Q000032:analysis; D009569:Nitric Oxide / Q000378:metabolism; D051304:Profilins / Q000032:analysis; D055333:Salivary Cystatins / Q000032:analysis; D012471:Salivary Proteins and Peptides / Q000032:analysis* / Q000378:metabolism; D013295:Streptococcus mutans / Q000378:metabolism*; D013682:Teichoic Acids / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000187:drug effects; D037521:Virulence Factors / Q000378:metabolism
D000199:Actins; C082965:CALML3 protein, human; C525171:CST1 protein, human; C525030:CST3 protein, human; C525173:CST4 protein, human; D002147:Calmodulin; D054357:Chemokine CXCL10; C485433:Cxcl10 protein, mouse; D055316:Cystatin C; D015891:Cystatins; D023082:Defensins; D006657:Histones; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; C495738:PFN1 protein, human; D051304:Profilins; D055333:Salivary Cystatins; D012471:Salivary Proteins and Peptides; D013682:Teichoic Acids; D051195:Toll-Like Receptor 2; D037521:Virulence Factors; C071378:CST5 protein, human; D009569:Nitric Oxide; C009900:lipoteichoic acid; D009113:Muramidase; C514728:lysozyme C, human
Streptococcus mutans; dental caries; lipoteichoic acid; salivary proteins