CALMODULIN ACTIVATED ADENYLATE CYCLASE OF B. PERTUSSIS

我们的目的是表征波氏杆菌腺苷酸环化酶 百日咳，克隆其基因，并利用该基因产生更多的 蛋白质比可以从细菌中回收。 我们发现该 酶比以前想象的更复杂。 我们的生化研究 使我们能够开发亲和程序， 酵素 我们最初的实验也确定， 钙调素是直接的，我们已经开发了筛选该基因的方法 基于腺苷酸环化酶的能力， 肽结合钙调素。 克隆序列将通过测序表征，然后进行操作 用于几个目的，包括在E.杆菌 产生特定的博德特氏菌腺苷酸环化酶突变体， 该基因在真核细胞中的表达。 物理表征后，解决了一些相互矛盾的结果 在文献中，我们将研究腺苷酸环化酶对 哺乳动物靶细胞。 最后，我们将利用基因的不寻常进化， 试图用它来长期诱导腺苷酸环化酶和cAMP， 哺乳动物细胞

Our objective is to characterize the adenylate cyclase of Bordetella pertussis, to clone its gene, and to use the gene to produce more of the protein than can be recovered from the bacterium. We have found that this enzyme is more complex than previously thought. Our biochemical studies allowed us to develop affinity procedures which we will use to purify this enzyme. Our initial experiments have also determined that binding of calmodulin is direct and we have developed methods to screen for the gene in recombinant DNA libraries based on the ability of the adenylate cyclase peptide to bind calmodulin. Cloned sequences will be characterized by sequencing and then manipulated for several purposes which include the production of enzyme in E. coli, the creation of specific Bordetella adenylate cyclase mutants, and the expression of the gene in eucaryotic cells. After physical characterization to resolve some of the conflicting results in the literature, we will study action of the adenylate cyclcase on mammalian target cells. Finally we will take advantage of the unusual evolution of the gene and attempt to use it for long term induction of adenylate cyclase and cAMP in mammalian cells.