Interaction of 7-bromoacetyl-7-desacetylforskolin with adenylate cyclase
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-; TRN: 86-031393
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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CYCLASES
ENZYME ACTIVITY
PROTEINS
BIOCHEMICAL REACTION KINETICS
BLOOD PLATELETS
BRAIN
CATTLE
CELL MEMBRANES
COVALENCE
MAN
MOLECULAR WEIGHT
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM
DOMESTIC ANIMALS
ENZYMES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LYASES
MAMMALS
MATERIALS
MEMBRANES
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
PRIMATES
REACTION KINETICS
RUMINANTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques