title: Calmodulin inhibits entry of Bordetella pertussis adenylate cyclase into animal cells.
abstract: Bordetella pertussis, the pathogen responsible for whooping cough, releases a soluble calmodulin-sensitive adenylate cyclase into its culture medium. Recently, Confer and Eaton [Confer, D., & Eaton, J. (1982) Science (Washington, D.C.) 217, 948-950], as well as Hanski and Farfel [Hanski, E., & Farfel, Z. (1985) J. Biol. Chem. 290, 5526-5536], have shown that crude extracts from B. pertussis containing adenylate cyclase activity cause elevations in intracellular cAMP when incubated with human neutrophils or lymphocytes. These investigators proposed that the bacterial enzyme enters animal cells and catalyzes the formation of cAMP from intracellular ATP. In this study, B. pertussis adenylate cyclase was purified to remove contaminating islet activating protein and examined for its effects on intracellular cAMP levels of human erythrocytes and N1E-115 mouse neuroblastoma cells. In both cases, the enzyme catalyzed the formation of intracellular cAMP. Addition of calmodulin to the adenylate cyclase preparations completely inhibited formation of intracellular cAMP catalyzed by the bacterial enzyme, indicating that cAMP was not synthesized extracellularly and then taken up by the cells. These experiments illustrate that the bacterial enzyme does enter animal cells and that the enzyme-calmodulin complex does not.
authors.forename: R L
authors.initials: RL
authors.lastname: Shattuck
authors.forename: D R
authors.initials: DR
authors.lastname: Storm
mesh_terms: D000262:Adenylyl Cyclases / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001886:Bordetella pertussis / Q000201:enzymology*; D001921:Brain; D002147:Calmodulin / Q000494:pharmacology*; D002417:Cattle; D002460:Cell Line; D000242:Cyclic AMP / Q000097:blood; D004789:Enzyme Activation; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D007700:Kinetics
chemical_list: D002147:Calmodulin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases