cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor.

Abstract

We have isolated and sequenced a cDNA encoding the human 132-adrenergic receptor. The deduced amino acid sequence (413 residues) is that of a protein containing seven clusters of hydrophobic amino acids suggestive of membrane-spanning domains. While the protein is 87% identical overall with the previously cloned hamster P2-adrenergic receptor, the most highly conserved regions are the putative transmembrane helices (95% identical) and cytoplasmic loops (93% identical), suggesting that these regions of the molecule harbor important functional domains. Several of the transmembrane helices also share lesser degrees of identity with comparable regions of select members of the opsin family of visual pigments. We have localized the gene for the P2-adrenergic receptor to q31-q32 on chromosome 5. This is the same position recently determined for the gene encoding the receptor for platelet-derived growth factor and is adjacent to that for the FMS protooncogene, which encodes the receptor for the macrophage colony-stimulating factor.Many hormones, neurotransmitters, and drugs influence cellular metabolic activities by stimulating the adenylate cyclase system, leading to the generation of the second messenger cAMP and activation of the cAMP-dependent protein kinase. The molecular components of this plasma membrane signaling system include specific receptors that bind ligands, the catalyst that converts ATP to cAMP, and guanine nucleotide regulatory or G proteins that functionally couple the receptors to the enzyme (1). The latter two components of the system have been purified and genes encoding several members of the "G protein family" have been cloned.Of the receptors that are coupled to adenylate cyclase the only one that has been characterized in any detail is the p-adrenergic receptor (,8AR). Two pharmacologically and physiologically distinct subtypes of this receptor, termed ,31AR and f32AR, are both membrane glycoproteins of Mr 64,000 (2). Very recently, we reported cloning of cDNA and the gene for the hamster f32AR (3). The deduced protein sequence indicated a protein of 418 amino acids, with seven clusters of hydrophobic amino acids likely representing membrane-spanning regions. This topology resembles that of the visual "light receptor" rhodopsin, which also possesses seven membrane-spanning domains (4-6).We now report the cloning and complete nucleotide sequence of the cDNA for the human ,82AR. While the receptor is highly similar to its hamster counterpart (87% of the amino acid residues are identical), significant regional differences in the extent of identity are noted. METHODScDNA Library Screening. The human placenta cDNA library was kindly provided by Evan Sadler (Washington University School of Medicine). The cDNA was prepared from term placenta poly(A)+ RNA and cloned in phage Xgtll. The library contains 5 x 106 independent recombinants. The A431 Xgtll library was prepared from poly(A)+ RNA from actively dividing A431 cells by methods previously described (3). It contains 1 x 106 independent reco...

Catechol- O -methyltransferase inhibition increases pain sensitivity through activation of both β2- and β3-adrenergic receptors

“…β 2 ARs are predominantly located on vascular, uterine, and airway smooth muscle (Dixon et al 1986) and mononuclear leukocytes in the periphery (Landmann 1992) as well as cerebellar and thalamic neurons (Nicholas et al 1996;Rainbow et al 1984) and glial cells (Salm and McCarthy 1992;Stone and Ariano 1989) in the central nervous system. Moderate levels of expression have also been observed in adipocytes (Kobilka et al 1987) and spinal dorsal horn neurons (Nicholson et al 2005). The contribution of β 2 ARs to enhanced pain sensitivity is in line with results from previous studies demonstrating that epinephrine activates β 2 ARs located on primary afferent nociceptors and produces a hyperalgesic state in rats (Aley et al 2001;Khasar et al 1999a;Khasar et al 1999b;Khasar et al 2003).…”

Electrophilic derivatives of purines as irreversible inhibitors of A1 adenosine receptors

“…In addition, chain lengths were varied to probe the architecture of the binding site. Lysyl residues, histidyl residues, α-amino groups, and sulfhydryl groups are known to be present in the primary sequence of other membrane-bound receptors (for example, β 2 -adrenergic receptors 19 ) in the vicinity of the extracellular binding site. Evidence suggestive of a free thiol group present on the A 1 receptor has been reported.…”

Pharmacogenetics of the β2-Adrenergic Receptor Gene

“…ADRβ2 is a small, intronless gene with one exon that encodes for a 413-amino acid, G-protein coupled receptor, the β2-adrenergic receptor. 28,29 ADRβ2 is an ideal candidate gene for the study of asthma genetics due to its location on chromosome 5q31: a region within the human genome consistently linked to asthma and related phenotypes (bronchial hyperresponsiveness (BHR), and serum IgE levels) through large family-based linkage studies in the Dutch, Caucasians, African Americans, and U.S. Hispanics. [30][31][32][33] The linkage to asthma susceptibility within this region of the human genome has also been shown to have a gene-environment interaction with passive smoking exposure.…”