title: Purification and characterization of a calmodulin-sensitive adenylate cyclase from Bordetella pertussis.
abstract: Bordetella pertussis, the bacterium responsible for whooping cough, releases a soluble, calmodulin-sensitive adenylate cyclase into its culture medium. B. pertussis mutants deficient in this enzyme are avirulent, indicating that the adenylate cyclase contributes to the pathogenesis of the disease. It has been proposed that B. pertussis adenylate cyclase may enter animal cells and increase intracellular adenosine cyclic 3',5'-phosphate (cAMP) levels. We have purified the enzyme extensively from culture medium using anion-exchange chromatography in the presence and absence of calmodulin and gel filtration chromatography. The enzyme was purified 1600-fold to a specific activity of 608 mumol of cAMP min-1 mg-1 and was free of islet activating protein. The molecular weight of the enzyme was 43 400 in the absence of calmodulin and 54 200 in the presence of calmodulin. The Km of the bacterial enzyme for adenosine 5'-triphosphate was 2.0 mM, whereas the Km of the calmodulin-sensitive adenylate cyclase from bovine brain was 0.07 mM. Although the enzyme was not purified to homogeneity, its turnover number of 27 000 min-1 is the highest documented for any adenylate cyclase preparation.
authors.forename: R L
authors.initials: RL
authors.lastname: Shattuck
authors.forename: D J
authors.initials: DJ
authors.lastname: Oldenburg
authors.forename: D R
authors.initials: DR
authors.lastname: Storm
mesh_terms: D000262:Adenylyl Cyclases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001886:Bordetella pertussis / Q000201:enzymology*; D001921:Brain; D002147:Calmodulin / Q000302:isolation & purification / Q000494:pharmacology*; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D007700:Kinetics
chemical_list: D002147:Calmodulin; D000262:Adenylyl Cyclases