title: Purification of bovine brain adenylyl cyclase with a novel derivative of forskolin: evidence for a high specific activity form of the enzyme.
abstract: An improved affinity support for the purification of adenylyl cyclase was prepared from 7-desacetyl-7-aminoethylaminocarbonyl forskolin. This analog allows convenient synthesis of an affinity matrix that is chemically stable, with-standing repeated use for up to two years, and efficient, yielding purifications of adenylyl cyclase from solubilized bovine brain membranes of 2,000-6,000 fold in a single step. Immunoblotting data suggest that the majority of the enzyme purified in this fashion differs from forms described previously. Since the specific activity of this preparation is substantially higher than that described in previous reports, it is possible that the purification described here selects, presumably on the basis of affinity for forskolin, for a form of adenylyl cyclase with higher specific activity than any described previously.
authors.affiliation: Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA.
authors.forename: M
authors.initials: M
authors.lastname: Moos
authors.forename: D I
authors.initials: DI
authors.lastname: Morris
authors.forename: J
authors.initials: J
authors.lastname: Robbins
authors.forename: L
authors.initials: L
authors.lastname: Appel
authors.forename: K B
authors.initials: KB
authors.lastname: Seamon
mesh_terms: D000262:Adenylyl Cyclases / Q000302:isolation & purification*; D000327:Adsorption; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D005576:Colforsin / Q000031:analogs & derivatives*; D004224:Diterpenes; D015394:Molecular Structure; D009994:Osmolar Concentration; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: C102849:7-desacetyl-7-aminoethylaminocarbonyl forskolin; D004224:Diterpenes; D005576:Colforsin; D000262:Adenylyl Cyclases