Streptococcus salivarius fimbriae are composed of a glycoprotein containing a repeated motif assembled into a filamentous nondissociable structure.
Streptococcus salivarius, a gram-positive bacterium found in the human oral cavity, expresses flexible peritrichous fimbriae. In this paper, we report purification and partial characterization of S. salivarius fimbriae. Fimbriae were extracted by shearing the cell surface of hyperfimbriated mutant A37 (a spontaneous mutant of S. salivarius ATCC 25975) with glass beads. Preliminary experiments showed that S. salivarius fimbriae did not dissociate when they were incubated at 100 degrees C in the presence of sodium dodecyl sulfate. This characteristic was used to separate them from other cell surface components by successive gel filtration chromatography procedures. Fimbriae with molecular masses ranging from 20 x 10(6) to 40 x 10(6) Da were purified. Examination of purified fimbriae by electron microscopy revealed the presence of filamentous structures up to 1 microm long and 3 to 4 nm in diameter. Biochemical studies of purified fimbriae and an amino acid sequence analysis of a fimbrial internal peptide revealed that S. salivarius fimbriae were composed of a glycoprotein assembled into a filamentous structure resistant to dissociation. The internal amino acid sequence was composed of a repeated motif of two amino acids alternating with two modified residues: A/X/T-E-Q-M/phi, where X represents a modified amino acid residue and phi represents a blank cycle. Immunolocalization experiments also revealed that the fimbriae were associated with a wheat germ agglutinin-reactive carbohydrate. Immunolabeling experiments with antifimbria polyclonal antibodies showed that antigenically related fimbria-like structures were expressed in two other human oral streptococcal species, Streptococcus mitis and Streptococcus constellatus.
Lévesque
C
C
Groupe de Recherche en Ecologie Buccale, Faculté de Médecine Dentaire, Université Laval, Quebec City, Quebec, Canada.
Vadeboncoeur
C
C
Chandad
F
F
Frenette
M
M
D000596:Amino Acids / Q000235:genetics; D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000648:ultrastructure; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D006358:Hot Temperature; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D009154:Mutation; D016624:Negative Staining; D012688:Septum Pellucidum; D012967:Sodium Dodecyl Sulfate; D013291:Streptococcus / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D000596:Amino Acids; D006023:Glycoproteins; D014909:Wheat Germ Agglutinins; D012967:Sodium Dodecyl Sulfate