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Salivary Receptors for the Proline-rich Protein-binding and Lectin-like Adhesins of Oral Actinomyces and Streptococci

Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, Building 30, Room 532, National Institutes of Health, Bethesda, MD 20892;

View larger version (64K): [in a new window]   Figure 1. Detection of salivary proteins and glycoproteins following separation of parotid (P) and SM-SL (S) saliva by SDS-PAGE. (A) Gels were silver-stained or transferred to nitrocellulose prior to the labeling of amino and sulfhydryl groups. Glycosylated proteins were labeled and detected on blots following oxidation of sialic acids with 1 mM sodium periodate or total carbohydrate with 10 mM sodium periodate (IO4–). (B) Individual blots were overlaid with biotinylated lectins, and bound lectin was detected with avidin-D alkaline phosphatase. The lectin probes used were LFA for terminal sialic acid, PNA for terminal Galß1-3GalNAc, LTA for L-fucose, and ConA for glucose and mannose. Blots remained untreated (UT) or were sialidase-treated (SIAL) prior to incubation with lectins. The locations of purified MG1, MG2, PRG, -amylase (AMY), and PRP-1 are indicated.

View larger version (48K): [in a new window]   Figure 2. Adhesion of biotinylated A. naeslundii parent and fimbriae-deficient mutant strains to: (A) nitrocellulose membranes spotted with 10 ng purified PRP-1 or PRG and 1 µg MG2 or asialo-MG2; or (B) blots of parotid (P) and SM-SL (S) secretions separated by SDS-PAGE. The presence or absence of type 1 and type 2 fimbriae on each strain is indicated in parentheses. Adherent bacteria were detected with avidin-D alkaline phosphatase. Adhesion assays were also performed in the presence of 2 mM EGTA where indicated. Blots remained untreated (UT) or were sialidase-treated (SIAL) prior to the addition of bacteria.

View larger version (38K): [in a new window]   Figure 3. Adhesion of biotinylated S. gordonii DL1, homologous mutant strain D102, S. gordonii M5, S. sanguis 10556, or S. gordonii 10558 to: (A) nitrocellulose membranes spotted with 10 ng purified PRP-1 or PRG and 1 µg MG2 or asialo-MG2; or (B) blots of parotid (P) and SM-SL (S) secretions separated by SDS-PAGE. Adherent bacteria were detected with avidin-D alkaline phosphatase. Adhesion assays were also performed in the presence of 2 mM EGTA where indicated. Blots remained untreated (UT) or were sialidase-treated (SIAL) prior to the addition of bacteria.