title: Association of adenylate cyclase with an actin-like protein in the human myometrium.
abstract: Regulation of muscle contraction by second messengers such as cAMP and regulation of the adenylate cyclase enzyme by the cytoskeleton have been previously described. However, the physical association of both effector and structural elements is still unknown. In this context, we have co-purified a human myometrial adenylate cyclase with an actin-like protein in a two-step purification protocol. The adenylate cyclase catalytic unit was solubilized with Lubrol-PX, submitted to anionic exchange chromatography and purified about 7-fold. The eluate was applied to a forskolin-agarose column obtaining an adenylate cyclase extract enriched 257-fold (enzymatic activity of 1390 pmol/30 min per mg protein) that co-eluted with a 74.6-kDa protein that possessed the 18-27 amino-acid fragment from the N-terminal region of human actin. Under non-reducing conditions, the apparent molecular weight of this protein decreased to 54 kDa, which has been previously described for arthrin. These results provide the first demonstration of the physical association of human myometrial adenylate cyclase with a cytoskeleton-related protein, supporting the hypothesis that adenylate cyclase is regulated by mechanical stimuli.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Alcalá, Alcalá de Henares, Spain.
authors.forename: A M
authors.initials: AM
authors.lastname: Bajo
authors.forename: J C
authors.initials: JC
authors.lastname: Prieto
authors.forename: P
authors.initials: P
authors.lastname: Valenzuela
authors.forename: P
authors.initials: P
authors.lastname: Martínez
authors.forename: C
authors.initials: C
authors.lastname: Menor
authors.forename: A
authors.initials: A
authors.lastname: Marina
authors.forename: J
authors.initials: J
authors.lastname: Vázquez
authors.forename: L G
authors.initials: LG
authors.lastname: Guijarro
mesh_terms: D000262:Adenylyl Cyclases / Q000187:drug effects / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D005576:Colforsin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D009215:Myometrium / Q000201:enzymology*
chemical_list: D005576:Colforsin; D000262:Adenylyl Cyclases