Open AccessPurification and characterization of a hemoglobin‐binding outer membrane protein of Prevotella intermedia
Author(s) -
Guan SuMin,
Nagata Hideki,
Maeda Kazuhiko,
Kuboniwa Masae,
Minamino Naoto,
Shizukuishi Satoshi
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09607.x
Subject(s) - hemoglobin , dissociation constant , hemeprotein , biochemistry , binding protein , western blot , chemistry , bacterial outer membrane , myoglobin , microbiology and biotechnology , hemin , protein g , biology , heme , enzyme , escherichia coli , antibody , receptor , immunology , gene
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 × 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.