Soluble low affinity adenosine A/sub 2/ binding site from human placenta: reconstitution and characteristics

Conference · Fri May 01 00:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6126878

Hutchison, K; Prasad, M; Fox, I H

The authors have developed a vesicle reconstitution technique that allows for rapid vacuum filtration assay, and have characterized the soluble A/sub 2/ site from placental membranes. The overall yield of reconstituted binding is 60%. Competition analysis of membranes and reconstituted vesicles yields identical agonist potency orders and affinities: N-ethylcarboxamidoadenosine (NECA) (Kd-330 nM)>2-chloroadenosine (Kd=1.7 ..mu..M) > L-phenylisopropyladenosine (Kd > 1 mM). Equilibrium binding to membranes and reconstituted vesicles of (/sup 3/H)-NECA, an adenosine agonist, was not reduced by guanine nulceotides. HPLC gel permeation chromatography of extracts from membranes preincubated with 5 mM MgCl/sub 2/ and 100 ..mu..M NECA revealed a peak of binding with kD of 0.07. Extracts prepared with either an antagonist or NECA and 100 ..mu..M guanylyl 5'-imidodiphosphate revealed a peak of binding with a kD of 0.09. These data suggest that the adenosine A/sub 2/ receptor retains its binding properties upon reconstitution and may couple to a guanine nucleotide regulatory protein.

• Research Organization: Univ. Hospitals, Ann Arbor, MI
• OSTI ID: 6126878
• Report Number(s): CONF-870644-; TRN: 87-033989
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ADENOSINE
RECEPTORS
BIOCHEMICAL REACTION KINETICS
AFFINITY
CELL MEMBRANES
LIQUID COLUMN CHROMATOGRAPHY
MAN
PLACENTA
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
CELL CONSTITUENTS
CHROMATOGRAPHY
FETAL MEMBRANES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
REACTION KINETICS
RIBOSIDES
SEPARATION PROCESSES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques