Open AccessPurification and characterization of Bordetella calmodulin‐like protein
Author(s) -
Nagai Masaaki,
Endoh Masahiko,
Danbara Hirofumi,
Nakase Yasukiyo
Publication year - 1994
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.1574-6968.1994.tb06696.x
Subject(s) - calmodulin , bordetella pertussis , bordetella bronchiseptica , bordetella , biochemistry , polyacrylamide gel electrophoresis , phosphodiesterase , biology , gel electrophoresis , microbiology and biotechnology , chemistry , bacteria , enzyme , genetics
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 Ca 2+ 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.