Synthetic peptides analogous to the fimbrillin sequence inhibit adherence of Porphyromonas gingivalis.
Fimbriae are important in the adherence of many bacterial species to the surfaces they eventually colonize. Porphyromonas (Bacteroides) gingivalis fimbriae appear to mediate adherence to oral epithelial cells and the pellicle-coated tooth surface. The role and contribution of fimbriae in the binding of P. gingivalis to hydroxyapatite (HAP) coated with saliva as a model for the pellicle-coated tooth surface were investigated. 3H-labeled P. gingivalis or the radioiodinated purified fimbriae were incubated with 2 mg of HAP beads coated with whole human saliva (sHAP) and layered on 100% Percoll to separate unbound from sHAP-bound components. The radioactivity of the washed beads was a measure of the bound components. The binding of P. gingivalis 2561 (381) cells and that of purified fimbriae were concentration dependent and saturable at approximately 10(8) cells and 40 micrograms of fimbriae added, respectively. The addition of fimbriae inhibited binding of P. gingivalis to sHAP beads by 65%, while the 75-kDa protein, which is another major surface component of P. gingivalis 2561, did not show significant inhibition, suggesting that the fimbriae are important in adherence. Encapsulated and sparsely fimbriated P. gingivalis W50 did not bind to sHAP beads. On the basis of the predicted sequence of the fimbrillin, a structural subunit of fimbriae, a series of peptides were synthesized and used to localize the active fimbrillin domains involved in P. gingivalis adherence to sHAP beads. Peptides from the carboxyl-terminal one-third of the fimbrillin strongly inhibited P. gingivalis binding to sHAP beads. Active residues within the sequence of inhibitory peptide 226-245 (peptide containing residues 226 to 245) and peptide 293-306 were identified by using smaller fragments prepared either by trypsin cleavage of the peptide 226-245 or by synthesis of smaller segments of peptide 293-306. Hemagglutinin activity, lectinlike binding, and ionic interaction did not seem to be involved in this binding since lysine, arginine, carbohydrates, and calcium ions failed to affect the binding of P. gingivalis. The observation that poly-L-lysine, bovine serum albumin, and defatted bovine serum albumin, even at high concentrations, only partially blocked the binding of P. gingivalis indicates that hydrophobic interactions are not the major forces involved in P. gingivalis binding to sHAP beads. Protease inhibitors such as EDTA, leupeptin, pepstatin, 1,10-phenanthroline, and phenylmethylsulfonyl fluoride did not interfere with the binding of P. gingivalis. However, the binding of P. gingivalis to trypsin- or chymotrypsin-pretreated sHAP beads was reduced.(ABSTRACT TRUNCATED AT 400 WORDS)
Department of Oral Biology, State University of New York, Buffalo 14214.
J Y
JY
Lee
H T
HT
Sojar
G S
GS
Bedi
R J
RJ
Genco
D000595:Amino Acid Sequence; D001422:Bacterial Adhesion / Q000187:drug effects / Q000502:physiology; D001426:Bacterial Proteins / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002241:Carbohydrates / Q000494:pharmacology; D002918:Chymotrypsin / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000502:physiology; D006388:Hemagglutinins / Q000502:physiology; D037102:Lectins / Q000502:physiology; D008239:Lysine / Q000494:pharmacology; D008969:Molecular Sequence Data; D011107:Polylysine / Q000494:pharmacology; D016966:Porphyromonas gingivalis / Q000472:pathogenicity*; D011480:Protease Inhibitors / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D001426:Bacterial Proteins; D002241:Carbohydrates; D006388:Hemagglutinins; D037102:Lectins; D011480:Protease Inhibitors; C063055:fimbrillin; D036781:Fimbriae Proteins; D011107:Polylysine; D012710:Serum Albumin, Bovine; D002918:Chymotrypsin; D014357:Trypsin; D008239:Lysine; D002118:Calcium