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Deacetylation of forskolin catalyzed by bovine brain membranes

Journal Article · · Biochem. Biophys. Res. Commun.; (United States)
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Radiolabeled forskolin, 7-(/sup 3/H-acetyl)-forskolin, was synthesized to explore interactions between forskolin and bovine brain membrane preparations. The radiolabeled derivative was chemically characterized, and found to be indistinquishable from unlabeled forskolin in its ability to stimulate bovine brain adenylate cyclase. Preliminary binding data demonstrated that binding of 7-(/sup 3/H-acetyl)-forskolin to membranes was concentration dependent. However, competition binding studies using a constant concentration of 7-(/sup 3/H-acetyl)-forskolin with increasing levels of unlabeled forskolin showed enhanced binding of the labeled derivative. This suggested that 7-(/sup 3/H-acetyl)-forskolin was degraded by membranes and protected by native forskolin. Incubation of forskolin with membranes and analysis of the products by thin layer chromatography and mass spectroscopy showed the formation of 7-desacetylforskolin. The deacetylation of forskolin was monitored by quantitating the release of (/sup 3/H)acetate from 7-(/sup 3/H-acetyl)-forskolin. The reaction was linear with time and protein concentration. These data illustrate that forskolin can be degraded by membranes and indicate that ligand binding studies using labeled forskolin and membrane preparations should be cautiously interpreted.

OSTI ID:
5809756
Journal Information:
Biochem. Biophys. Res. Commun.; (United States), Journal Name: Biochem. Biophys. Res. Commun.; (United States) Vol. 1; ISSN BBRCA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
ACETYLATION
ACYLATION
ANIMALS
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CARBOXYLIC ACID SALTS
CATTLE
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
CHEMICAL REACTIONS
CHROMATOGRAPHY
CPB
CYCLASES
DOMESTIC ANIMALS
ENZYMES
IN VITRO
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LIGANDS
LYASES
MAMMALS
MASS SPECTROSCOPY
MEMBRANES
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RATS
REACTION KINETICS
RODENTS
RUMINANTS
SEPARATION PROCESSES
SPECTROSCOPY
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES