Cloning, pharmacological characterization, and polymorphism screening of the guinea pig beta(2)-adrenoceptor.
In asthma, beta(2)-adrenoceptor agonist responsiveness has been associated with Arg16Gly and Gln27Glu polymorphisms of the beta(2)-adrenoceptor. Since the guinea pig is extensively used as an animal model for asthma, we investigated the occurrence of possible polymorphism of the guinea pig beta(2)-adrenoceptor. The guinea pig beta(2)-adrenoceptor coding region was amplified by sequence homology-based cloning. Homology of the translated protein with the human beta(2)-adrenoceptor was 88% with Ala at position 16 and Glu at position 27. Radioligand binding and cAMP- accumulation experiments of Chinese hamster ovary (CHO) cells transfected with the guinea pig beta(2)-adrenoceptor revealed a homogeneous population of functional receptors. Five degenerate single nucleotide polymorphisms were found within the beta(2)-adrenoceptor coding region of outbred Dunkin Hartley guinea pigs, at residues 354, 453, 483, 534 and 642. In conclusion, we have cloned the guinea pig beta(2)-adrenoceptor, which shows to be functional upon expression in a recombinant system and contains five single nucleotide polymorphisms dissimilar to human polymorphisms.
Oostendorp
Jaap
J
Department of Molecular Pharmacology, University Centre for Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands. j.oostendorp@farm.rug.nl
Meurs
Herman
H
Adriaan Nelemans
S
S
Zaagsma
Johan
J
Kauffman
Henk F
HF
Postma
Dirkje S
DS
Boddeke
Hendrikus W G M
HW
Biber
Knut
K
D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D006168:Guinea Pigs; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D011869:Radioligand Assay; D018343:Receptors, Adrenergic, beta-2 / Q000187:drug effects* / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013999:Timolol / Q000494:pharmacology; D014162:Transfection
D000318:Adrenergic beta-Agonists; D000319:Adrenergic beta-Antagonists; D018343:Receptors, Adrenergic, beta-2; D013999:Timolol; D007545:Isoproterenol