title: Purification and characterization of Bordetella calmodulin-like protein.
abstract: Bordetella calmodulin-like protein was purified from culture supernatant fluid of B. pertussis, B. parapertussis and B. bronchiseptica by successive chromatography on hydroxyapatite, Toyopearl HW-50F and QAE-Toyopearl 550C columns. The purified calmodulin-like protein appeared to be homogeneous by SDS-polyacrylamide gel electrophoresis. The apparent molecular mass of calmodulin-like protein on SDS-polyacrylamide gel electrophoresis was 10 kDa, which was smaller than bovine brain calmodulin (17 kDa). The purified calmodulin-like protein activated both Bordetella adenylate cyclase and mammalian phosphodiesterase in a Ca(2+)-dependent manner. This activation was inhibited by calmodulin antagonists. The calmodulin-like protein, like calmodulin, was retained by a hydrophobic resin in the presence of Ca2+ and eluted by the addition of EDTA. These results indicated that the Bordetella calmodulin-like protein is closely related to calmodulin. As a putative calmodulin the extracellular calmodulin may be involved in Bordetella pathogenesis.
authors.affiliation: Department of Microbiology, School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Nagai
authors.forename: M
authors.initials: M
authors.lastname: Endoh
authors.forename: H
authors.initials: H
authors.lastname: Danbara
authors.forename: Y
authors.initials: Y
authors.lastname: Nakase
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D001884:Bordetella / Q000737:chemistry* / Q000254:growth & development; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D002147:Calmodulin / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D007526:Isoelectric Point; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000032:analysis
chemical_list: D001426:Bacterial Proteins; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D010727:Phosphoric Diester Hydrolases; D000262:Adenylyl Cyclases