title: Binding of hemoglobin by Porphyromonas gingivalis.
abstract: In this study, we investigated whether Porphyromonas gingivalis can bind hemoglobin as an initial step in the acquisition of heme from hemoglobin. The binding of human hemoglobin by P. gingivalis cells was determined using [3H]hemoglobin. Hemoglobin binding occurred rapidly, reversibly and specifically. A Scatchard analysis of the binding data generated a linear plot, indicating a single population of binding proteins. The apparent Kd was 1.0 +/- 0.19 x 10(-6) M and there were 3.2 +/- 0.76 x 10(4) binding sites per cell. Hemoglobin binding was inhibited by unlabeled human hemoglobin but not by hemin and protoporphyrin IX. The binding was only partially inhibited by human serum albumin, transferrin, lactoferrin, catalase and cytochrome c. These results suggest that the ligand recognized by the binding protein may not be the heme moiety. The binding of hemoglobin considerably increased when the organisms were grown under hemin-limited conditions. Hemoglobin bound to outer membrane proteins extracted from P. gingivalis cells on a dot blot binding assay and binding ability was lost after heating bacterial proteins. These results suggest that P. gingivalis cells interact with human hemoglobin through specific binding sites on their surfaces as a preliminary step in iron acquisition.
authors.affiliation: Department of Preventive Dentistry, Osaka University Faculty of Dentistry, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Amano
authors.forename: M
authors.initials: M
authors.lastname: Kuboniwa
authors.forename: K
authors.initials: K
authors.lastname: Kataoka
authors.forename: K
authors.initials: K
authors.lastname: Tazaki
authors.forename: E
authors.initials: E
authors.lastname: Inoshita
authors.forename: H
authors.initials: H
authors.lastname: Nagata
authors.forename: H
authors.initials: H
authors.lastname: Tamagawa
authors.forename: S
authors.initials: S
authors.lastname: Shizukuishi
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001665:Binding Sites; D006427:Hemin / Q000502:physiology; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D016966:Porphyromonas gingivalis / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D006454:Hemoglobins; D011506:Proteins; D006427:Hemin