Abstract

Streptococcus mutans, the cause of dental caries, contains antigens crossreactive with human cardiac and skeletal muscle. This finding requires caution in development of anti-caries vaccines and demands intensive investigations of the immunochemistry of walls and membranes of this oral inhabitant. Recent studies of the S. mutans BHT membrane have revealed it to be the site of heart cross-reactive antigens (HRA). Further studies of the biology and immunochemistry of S. mutans membranes, therefore, appear warranted. Membranes of BHT will be analyzed by crossed immunoelectrophoresis (CIE)-zymogram techniques for adenosine triphosphatase, polynucleotide phosphorylase, aminopeptidase and proteases. Physical properties of enzymes will be determined by nitrocellulose blotting of SDS-PAGE and isoelectric focusing gels followed by zymogram development. Penicillin-binding proteins will be studied in membranes from cells at various growth stages using 14C-benzylpenicillin-binding. The permease, Enzyme II, of the PEP-dependent phosphotransferase system (PTS) will be detected using radioiodinated soluble PTS components as probes to develop blots of two-dimensional gels. Studies of induced vs non-induced cells and PTS-mutants will allow identifications of specific permeases. HRA's in detergent extracts of membranes will be routinely identified by immunoblotting. Purification of membrane-bound or cell-free HRA will be by immunoaffinity chromatography employing monoclonal antibodies (MAb's). Hybridomas will be selected on their abilities to produce MAb's reactive with S. mutans and human heart. Purified HRA's will be chemically analyzed for comparison with wall-localized HRA's. Also, purified antigens will be studied as adherence factors by reacting with heart tissue followed by indirect immunofluorescence and indirect RIA. The further occurrence of membrane HRA's similar to that of BHT will be investigated by immunoblot analyses of membranes from other S. mutans serotypes, other viridans streptococci, and fresh isolates of S. pyogenes from rheumatics. Mab's to BHT HRA will be used in this survey. Finally, the Mab's will be used to develop immunoblots of human heart tissue extracts to confirm the tissue-localization of antigens with identical determinants. These will be compared with immunoblots developed with anti-S. pyogenes serum.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
7R01DE008007-01
Application #
3221787
Study Section
Oral Biology and Medicine Study Section (OBM)
Project Start
1986-03-01
Project End
1988-02-29
Budget Start
1986-03-01
Budget End
1987-02-28
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Florida
Department
Type
Schools of Dentistry/Oral Hygn
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611

  Publications

Binepal, Gursonika; Gill, Kamal; Crowley, Paula et al. (2016) Trk2 Potassium Transport System in Streptococcus mutans and Its Role in Potassium Homeostasis, Biofilm Formation, and Stress Tolerance. J Bacteriol 198:1087-100
Crowley, P J; Brady, L J (2016) Evaluation of the effects of Streptococcus mutans chaperones and protein secretion machinery components on cell surface protein biogenesis, competence, and mutacin production. Mol Oral Microbiol 31:59-77
Tang, Wenxing; Bhatt, Avni; Smith, Adam N et al. (2016) Specific binding of a naturally occurring amyloidogenic fragment of Streptococcus mutans adhesin P1 to intact P1 on the cell surface characterized by solid state NMR spectroscopy. J Biomol NMR 64:153-64
Sullan, Ruby May A; Li, James K; Crowley, Paula J et al. (2015) Binding forces of Streptococcus mutans P1 adhesin. ACS Nano 9:1448-60
Heim, Kyle P; Sullan, Ruby May A; Crowley, Paula J et al. (2015) Identification of a supramolecular functional architecture of Streptococcus mutans adhesin P1 on the bacterial cell surface. J Biol Chem 290:9002-19
Lewis, N E; Brady, L J (2015) Breaking the bacterial protein targeting and translocation model: oral organisms as a case in point. Mol Oral Microbiol 30:186-97
Williams, Matthew L; Crowley, Paula J; Hasona, Adnan et al. (2014) YlxM is a newly identified accessory protein that influences the function of signal recognition particle pathway components in Streptococcus mutans. J Bacteriol 196:2043-52
Liao, Sumei; Klein, Marlise I; Heim, Kyle P et al. (2014) Streptococcus mutans extracellular DNA is upregulated during growth in biofilms, actively released via membrane vesicles, and influenced by components of the protein secretion machinery. J Bacteriol 196:2355-66
Heim, Kyle P; Crowley, Paula J; Long, Joanna R et al. (2014) An intramolecular lock facilitates folding and stabilizes the tertiary structure of Streptococcus mutans adhesin P1. Proc Natl Acad Sci U S A 111:15746-51
Heim, Kyle P; Crowley, Paula J; Brady, L Jeannine (2013) An intramolecular interaction involving the N terminus of a streptococcal adhesin affects its conformation and adhesive function. J Biol Chem 288:13762-74

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