Abstract

One goal of this program is to prepare irreversible inhibitors of the cAMP-specific and of the calmodulin-sensitive forms of cyclic nucleotide phosphodiesterase. Affinity probes will be used to determine part of the amino acid composition of the hydrolytic site of the calmodulin-sensitive phosphodiesterase. The relationship between irreversible inhibition in intact porcine coronary arteries and the intracellular levels of cyclic AMP and cyclic GMP will be determined in order to establish which form of the enzyme is mainly responsible for the regulation of each nucleotide. In a second aspect of this program, we will attempt to photoaffinity label and biochemically characterize the A2-Adenosine receptor. We have developed potent and relatively selective inhibitors of the two subtypes of adenosine receptor and, based on the structure activity relationships of these inhibitors, we will also design and prepare high affinity ligands to use in characterization of the A1 and the A2-adenosine receptors.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM021220-11
Application #
3270339
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1978-09-01
Project End
1992-03-31
Budget Start
1987-04-01
Budget End
1988-03-31
Support Year
11
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
Vanderbilt University Medical Center
Department
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37203

  Publications

Kennedy, A P; Mangum, K C; Linden, J et al. (1996) Covalent modification of transmembrane span III of the A1 adenosine receptor with an antagonist photoaffinity probe. Mol Pharmacol 50:789-98
Saunders, C; Keefer, J R; Kennedy, A P et al. (1996) Receptors coupled to pertussis toxin-sensitive G-proteins traffic to opposite surfaces in Madin-Darby canine kidney cells. A1 adenosine receptors achieve apical and alpha 2A adrenergic receptors achieve basolateral localization. J Biol Chem 271:995-1002
Piersen, C E; True, C D; Wells, J N (1994) A carboxyl-terminally truncated mutant and nonglycosylated A2a adenosine receptors retain ligand binding. Mol Pharmacol 45:861-70
Piersen, C E; True, C D; Wells, J N (1994) 125I-2-[4-[2-[2-[(4-azidophenyl)methylcarbonylamino] ethylaminocarbonyl]ethyl]phenyl] ethylamino-5'-N-ethylcarboxamidoadenosine labels transmembrane span V of the A2a adenosine receptor. Mol Pharmacol 45:871-7
Pong, A S; Wells, J N (1994) Proteolysis of a membrane-bound protein in polyacrylamide gel slices using freeze-pulverization and gel electrophoresis techniques. Anal Biochem 217:163-5
Caruthers, M H; Beaton, G; Cummins, L et al. (1991) Synthesis and biochemical studies of dithioate DNA. Ciba Found Symp 158:158-66;discussion 166-8
Ahn, H S; Foster, M; Foster, C et al. (1991) Evidence for essential histidine and cysteine residues in calcium/calmodulin-sensitive cyclic nucleotide phosphodiesterase. Biochemistry 30:6754-60
Kuan, C J; Wells, J N; Jackson, E K (1990) Endogenous adenosine restrains renin release in conscious rats. Circ Res 66:637-46
Katsushima, T; Nieves, L; Wells, J N (1990) Structure-activity relationships of 8-cycloalkyl-1,3-dipropylxanthines as antagonists of adenosine receptors. J Med Chem 33:1906-10
Zhang, Y; Wells, J N (1990) The effects of chronic caffeine administration on peripheral adenosine receptors. J Pharmacol Exp Ther 254:757-63

Showing the most recent 10 out of 20 publications