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Interaction of 7-bromoacetyl-7-desacetylforskolin with adenylate cyclase

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5234201
; ;

7-Bromoacetyl-7-desacetylforskolin (BrAcFk) and the 12-tritio derivative (/sup 3/H-BrAckFk) were synthesized as alkylating analogs of forskolin. BrAcFk stimulated adenylate cyclase in human platelet and bovine brain membranes with an EC50 of 50..mu..M and inhibited /sup 3/H-forskolin binding to these membranes with a K/sub i/ of 300 nM. /sup 3/H-forskolin binding was decreased in membranes pretreated for 20 min with 10 ..mu..M BrAcFk. The i,9-dideoxy derivative of BrAcFk did not activate adenylate cyclase or inhibit /sup 3/H-forskolin binding. Proteins labelled by BrAcFk in solubilized preparations from bovine brain and human platelets were identified by fluorography of SDS gels. The two predominant bands labelled in the low and high molecular weight regions had molecular weights of 50,000 and 135,000 daltons respectively. The 135,000 dalton band identified by fluorography coeluted with adenylate cyclase activity on a Dupont GF450 column and has a molecular weight identical to that of the catalytic subunit determined by silver staining of SDS gels. These results suggest that BrAcFk can react covalently with the catalytic subunit of adenylate cyclase.

Research Organization:
Center for Drugs and Biologics/FDA, Bethesda, MD
OSTI ID:
5234201
Report Number(s):
CONF-8606151-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY
BODY FLUIDS
BRAIN
CATTLE
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
COVALENCE
CYCLASES
DOMESTIC ANIMALS
ENZYME ACTIVITY
ENZYMES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LYASES
MAMMALS
MAN
MATERIALS
MEMBRANES
MOLECULAR WEIGHT
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
PRIMATES
PROTEINS
REACTION KINETICS
RUMINANTS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES