Human lactoferrin binding to Porphyromonas gingivalis, Prevotella intermedia and Prevotella melaninogenica.
Human isolates of Porphyromonas gingivalis (n = 16), Prevotella intermedia n = 82) and Prevotella melaninogenica (n = 18) from diseased periodontal pockets were examined for interaction with human lactoferrin (HLf) in a standardized 125I-labeled protein binding assay. The highest HLf binding was found in P. intermedia strains, followed by P. gingivalis and P. melaninogenica. Further characterization of the interaction was performed with 1 representative strain from each species. HLf binding to P. gingivalis reached a saturation instantly and was optimal at pH 5.0-6.5. The corresponding values for P. melaninogenica were 90 min and pH 3.0-5.5. The HLf binding to the 2 strains seem to be nonspecific. In contrast, P. intermedia demonstrated specific binding, and a time-saturability within 60 min with an optimal uptake at pH 6.0-7.5. Scatchard analysis implied 45,000 receptors per cell with an affinity constant of 5.5 x 10(-7) M on P. intermedia strain 4H. The binding capacity in all 3 strains was affected by the culture medium. HLf binding components in these strains were susceptible to heat or proteases. Binding was eliminated in P. gingivalis and was enhanced in P. intermedia and P. melaninogenica by periodate treatment. Unlabeled HLf or bovine lactoferrin effectively displaced labeled HLf binding. Various proteins and carbohydrates did not inhibit HLf binding. Our data suggest that HLf binds to these periodontitis-associated species and that this mechanism is distinct from the previously known ligand interactions in oral bacteria.
Kalfas
S
S
University of Lund, Sweden.
Andersson
M
M
Edwardsson
S
S
Forsgren
A
A
Naidu
A S
AS
D000209:Acute-Phase Proteins / Q000378:metabolism*; D001439:Bacteroides / Q000378:metabolism*; D001667:Binding, Competitive; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism*; D010518:Periodontitis / Q000382:microbiology*; D016966:Porphyromonas gingivalis / Q000378:metabolism; D001443:Prevotella melaninogenica / Q000378:metabolism; D011485:Protein Binding; D012471:Salivary Proteins and Peptides / Q000378:metabolism*
D000209:Acute-Phase Proteins; D012471:Salivary Proteins and Peptides; D007781:Lactoferrin