Cloning and functional characterization of a human A1 adenosine receptor.
A human brain hippocampus cDNA library was screened by hybridization with a dog A1 adenosine receptor cDNA probe. Sequencing of the resulting clones identified a 978 residue open reading frame encoding a 326 amino acid polypeptide showing 95.7% similarity with the dog A1 adenosine receptor. Individual clones of stably transfected CHO cells expressing the human A1 receptor were obtained and tested for their response to the A1 agonist CPA [N6-cyclopentyladenosine] in the presence of forskolin. One clone was further characterized with respect to membrane binding of various adenosine agonists and antagonists. The rank order of affinities observed was typical of an A1 adenosine receptor. A Kd value of 2.28 nM was determined using [3H]DPCPX [dipropylcyclopentyl-xanthine], an A1 selective antagonist.
Libert
F
F
Institut de Recherche Interdisciplinaire, Faculte de Medecine, Universite Libre de Bruxelles, Belgium.
Van Sande
J
J
Lefort
A
A
Czernilofsky
A
A
Dumont
J E
JE
Vassart
G
G
Ensinger
H A
HA
Mendla
K D
KD
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D004285:Dogs; D006624:Hippocampus / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011983:Receptors, Purinergic / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014970:Xanthines / Q000378:metabolism
D015342:DNA Probes; D011983:Receptors, Purinergic; D014970:Xanthines; D005576:Colforsin; C048599:N(6)-cyclopentyladenosine; D004247:DNA; C051360:1,3-dipropyl-8-cyclopentylxanthine; D000241:Adenosine