Cloning and expression of a bovine adenosine A<sub>1</sub> receptor cDNA

Tucker, Amy L.; Linden, Joel; Robeva, Anna; D'Angelo, Drew D.; Lynch, Kevin R.

doi:10.1016/0014-5793(92)80338-h

Cited by 48 publications

(12 citation statements)

References 19 publications

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“…In the LLC-PKI cells, the A 1 AdoR migrated consistently between 38 and 42 kDa. We presume that the difference in migration of specific A 1 AdoR photolabeling in MDCKII cells (estimated at 43-45 kDa) versus LLC-PKI cells is due to different glycosylation in the two cellular backgrounds, since treatment of the A 1 AdoR in LLC-PKI cells with endoglycosidase F reduces the apparent molecular mass of the photolabeled A 1 AdoR to ϳ36 kDa (data not shown), a value consistent with the mass of the amino acids encoded by the A 1 AdoR cDNA and that previously reported (14,15). The finding that the A 1 AdoR is similarly enriched on the apical surface following expression in MDCKII and LLC-PKI cells is consistent with the interpretation that the polarization observed is a consequence of the A 1 AdoR structure and its interaction with shared renal epithelial trafficking mechanisms.…”

Section: Resultssupporting

confidence: 77%

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Saunders

Keefer

Kennedy

et al. 1996

Journal of Biological Chemistry

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Section: Resultssupporting

confidence: 77%

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Saunders

Keefer

Kennedy

et al. 1996

Journal of Biological Chemistry

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“…Note that increasing the G protein concentration in the reconstitution mix from 3 to 40 nM produces a progressively larger fraction of adenosine receptors in the high affinity conformation. These curves demonstrate the very high affinity expected of the bovine receptor for the 125 I-ABA ligand (33). Based on the data in Fig.…”

Section: Resultsmentioning

confidence: 99%

Role of the Prenyl Group on the G Protein γ Subunit in Coupling Trimeric G Proteins to A1 Adenosine Receptors

Yasuda

Lindorfer

Woodfork

et al. 1996

Journal of Biological Chemistry

102

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The coupling of receptors to heterotrimeric G proteins is determined by interactions between the receptor and the G protein ␣ subunits and by the composition of the ␤␥ dimers. To determine the role of the ␥ subunit prenyl modification in this interaction, the CaaX motifs in the ␥ 1 and ␥ 2 subunits were altered to direct modification with different prenyl groups, recombinant ␤␥ dimers expressed in the baculovirus/Sf9 insect cell system, and the dimers purified. The activity of the ␤␥ dimers was compared in two assays: formation of the high affinity agonist binding conformation of the A1 adenosine receptor and receptor-catalyzed exchange of GDP for GTP on the ␣ subunit. The ␤ 1 ␥ 1 dimer (modified with farnesyl) was significantly less effective than ␤ 1 ␥ 2 (modified with geranylgeranyl) in either assay. The ␤ 1 ␥ 1 -S74L dimer (modified with geranylgeranyl) was nearly as effective as ␤ 1 ␥ 2 in either assay. The ␤ 1 ␥ 2 -L71S dimer (modified with farnesyl) was significantly less active than ␤ 1 ␥ 2 . Using 125 I-labeled ␤␥ subunits, it was determined that native and altered ␤␥ dimers reconstituted equally well into Sf9 membranes containing A1 adenosine receptors. These data suggest that the prenyl group on the ␥ subunit is an important determinant of the interaction between receptors and G protein ␥ subunits.The membranes of all cells contain multiple receptors and transmembrane signaling systems that regulate cell function (1-5). One major unsolved problem in cell signaling is understanding how a cell selects its response to a hormone or growth factor given the possibilities available. The signaling mechanism used by receptors coupled to the heterotrimeric G proteins 1 provides an excellent example of the complexities. Current evidence suggests that specificity is determined at many levels in this pathway. In addition to the selectivity provided by the receptor itself, the interaction between the intracellular loops of the receptors and the ␣␤␥ subunits in the heterotrimer is a major determinant of specificity (6, 7). Interestingly, some receptors couple selectively to certain G protein ␣ subunits. For example, the ␤-adrenergic receptor couples primarily to members of the G s ␣ family (1, 6) and rhodopsin to the G t ␣ subunit (2). Other receptors, such as the angiotensin AT 1 or muscarinic receptors couple to multiple ␣ subunits including members of the G q and G i class of ␣ subunits (6, 8), leading to activation of multiple signaling networks by a given ligand (5).Although the receptor and the ␣ subunit provide one determinant of selectivity, the ␤␥ subunit is clearly required for the receptor to couple to the ␣ subunit (9, 10). Thus, a third level of selectivity may be provided by the type of ␤␥ subunit used to form the receptor-␣␤␥ complex. In addition, since the ␤␥ dimer can activate effectors directly (11-13), the diversity of these proteins may contribute to the specificity of signaling. Two lines of experimental evidence support this conclusion. Using recombinant ␤␥ subunits purified from Sf9 insect cel...

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“…A few years ago it was realized that two putative G-protein coupled receptors (RDC7 and RDC8, cloned from dog thyroid, Libert et al 1989) were identical with two types of adenosine receptors, Al and AZA, respectively (Maenhaut et al 1990;Libert et al 1991). Later, similar receptors were cloned from other species (Mahan et al 1991;Tucker et al 1992;Fink et al 1992). Even more recently two other receptor types, corresponding to the previously recognized A~B subtype (Stahle et al 1992) and to a novel subtype called A3 (Meyerhof et al 1991;Zhou et al 1992) were cloned.…”

Section: Adenosine Receptor Signallingmentioning

confidence: 98%

Purinoceptors in the Nervous System ^*

Fredholm

1995

Pharmacology & Toxicology

167

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The purine nucleoside adenosine and the purine nucleotide ATP play different roles in the nervous system. Adenosine acts on a family of G protein coupled receptors, collectively called adenosine receptors or P1 purinoceptors. Four members of this family have been cloned and pharmacologically characterized: A1, A2A, A2B and A3. Their distribution, pharmacology and biological roles are briefly discussed. In particular, the evidence that adenosine acting at A1 receptors regulates the release of several neurotransmitters and that adenosine acting at A2A receptors modulates dopaminergic transmission is summarized. ATP acts on receptors called P2 purinoceptors, which appear to fall into at least two main families--G protein coupled receptors and intrinsic ion channels. Their subclassification is becoming clearer as receptors are cloned and new selective agonists and/or antagonists are becoming available. There is an interesting potential for development of drugs targeted at purines or their receptors.

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Cloning and expression of a bovine adenosine A₁ receptor cDNA

Cited by 48 publications

References 19 publications

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Role of the Prenyl Group on the G Protein γ Subunit in Coupling Trimeric G Proteins to A1 Adenosine Receptors

Purinoceptors in the Nervous System ^*

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Cloning and expression of a bovine adenosine A1 receptor cDNA

Cited by 48 publications

References 19 publications

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Receptors Coupled to Pertussis Toxin-sensitive G-proteins Traffic to Opposite Surfaces in Madin-Darby Canine Kidney Cells

Role of the Prenyl Group on the G Protein γ Subunit in Coupling Trimeric G Proteins to A1 Adenosine Receptors

Purinoceptors in the Nervous System *

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Product

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About

Cloning and expression of a bovine adenosine A₁ receptor cDNA

Purinoceptors in the Nervous System ^*