title: Purification and characterization of a hemoglobin-binding outer membrane protein of Prevotella intermedia.
abstract: An outer membrane hemoglobin-binding protein of Prevotella intermedia was identified and purified in the present study. Hemoglobin-binding protein was purified via a series of column chromatographic methods. The molecular mass of the purified protein, which was approximately 60 kDa, was determined by SDS-PAGE. Hemoglobin binding of the protein was examined by Western blot and dot blot assays. Hemoglobin-binding activity was pH dependent; the strongest binding activity occurred at pH 5.0. Globin greatly decreased the binding activity, whereas transferrin, cytochrome C, and catalase exerted no effect. Dissociation constant between the 60-kDa protein and hemoglobin was 1.48 x 10(-8) M as measured by surface plasmon resonance. These results suggest that P. intermedia binds hemoglobin specifically through the 60-kDa outer membrane protein.
authors.affiliation: Department of Preventive Dentistry, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Su-Min
authors.initials: SM
authors.lastname: Guan
authors.forename: Hideki
authors.initials: H
authors.lastname: Nagata
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Maeda
authors.forename: Masae
authors.initials: M
authors.lastname: Kuboniwa
authors.forename: Naoto
authors.initials: N
authors.lastname: Minamino
authors.forename: Satoshi
authors.initials: S
authors.lastname: Shizukuishi
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D018713:Prevotella intermedia / Q000378:metabolism*; D020349:Surface Plasmon Resonance
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D006454:Hemoglobins; C017206:hemoglobin-binding protein, bacteria; D007501:Iron