title: Interaction of the two cytosolic domains of mammalian adenylyl cyclase.
abstract: Adenylyl cyclase activity can be reconstituted by simple mixture of the two cytosolic domains of the enzyme after their independent synthesis in Escherichia coli. We have synthesized and purified the C1a domain of type I adenylyl cyclase and the C2 domain of the type II enzyme to assess their interactions with each other and with the activators Gsalpha and forskolin. In the absence of an activator, the fragments associate with low affinity and display low catalytic activity. This basal activity can be stimulated more than 100-fold by either forskolin or activated Gsalpha. Further, the addition of these activators increases the apparent affinity of the fragments for each other. Stimulation of catalysis by Gsalpha and forskolin is synergistic. These data suggest a model wherein either Gsalpha or forskolin enhances association of the other activator with adenylyl cyclase, as well as facilitating the interaction between the C1 and C2 domains of the enzyme.
authors.affiliation: Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Whisnant
authors.forename: A G
authors.initials: AG
authors.lastname: Gilman
authors.forename: C W
authors.initials: CW
authors.lastname: Dessauer
mesh_terms: D000262:Adenylyl Cyclases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology*; D017931:DNA Primers; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D019204:GTP-Binding Proteins / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011485:Protein Binding
chemical_list: D017931:DNA Primers; D005576:Colforsin; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases