Agonist photoaffinity labeling of A1 adenosine receptors: persistent activation reveals spare receptors.

Abstract:

:This study describes experiments investigating the mechanism of activation of A1 adenosine receptors. Isolated rat fat cells were used as a cellular model. The A1 receptors of these cells were covalently labeled with the agonist photoaffinity label R-2-azido-N6-p-hydroxyphenylisopropyladenosine. The covalent incorporation of the label into the binding subunit of the receptor was verified by demonstration of specific labeling of a peptide with Mr = 35,000 by the radioiodinated label. Such covalent labeling followed by removal of label not covalently bound led to a concentration-dependent reduction of cellular cAMP levels. This persistent effect of covalent labeling occurred with an IC50 value of 9 nM compared to an IC50 value of 0.9 nM for the direct reduction of cAMP levels by the label. The affinity of the label was determined in binding experiments. The Ki value of 19 nM was about 20 times higher than the corresponding IC50 value of cAMP reduction. Finally, the comparison between covalent binding and its effects suggests that covalently labeled receptors were fully activated. The data are interpreted as evidence for a receptor activation according to the occupancy theory. The analysis of the various concentration-response curves reveals the presence of spare receptors, which can be demonstrated by the method of agonist photoaffinity labeling.

journal_name

Mol Pharmacol

journal_title

Molecular pharmacology

authors

Lohse MJ,Klotz KN,Schwabe U

subject

Has Abstract

pub_date

1986-10-01 00:00:00

pages

403-9

issue

4

eissn

0026-895X

issn

1521-0111

journal_volume

30

pub_type

杂志文章
  • State-dependent etomidate occupancy of its allosteric agonist sites measured in a cysteine-substituted GABAA receptor.

    abstract::A central axiom of ligand-receptor theory is that agonists bind more tightly to active than to inactive receptors. However, measuring agonist affinity in inactive receptors is confounded by concomitant activation. We identified a cysteine substituted mutant γ-aminobutyric acid type A (GABAA) receptor with unique chara...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.112.084558

    authors: Stewart DS,Hotta M,Desai R,Forman SA

    更新日期:2013-06-01 00:00:00

  • Activation of retinoic acid receptors by dihydroretinoids.

    abstract::Vitamin A-derived metabolites act as ligands for nuclear receptors controlling the expression of a number of genes. Stereospecific saturation of the C(13)-C(14) double bond of all-trans-retinol by the enzyme, retinol saturase (RetSat), leads to the production of (R)-all-trans-13,14-dihydroretinol. In liver and adipose...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.109.060038

    authors: Moise AR,Alvarez S,Domínguez M,Alvarez R,Golczak M,Lobo GP,von Lintig J,de Lera AR,Palczewski K

    更新日期:2009-12-01 00:00:00

  • PAR1 and PAR2 couple to overlapping and distinct sets of G proteins and linked signaling pathways to differentially regulate cell physiology.

    abstract::The protease-activated receptors (PAR1 and PAR2) are unusual G protein-coupled receptors that are activated by distinct serine proteases and are coexpressed in many different cell types. Limited recent evidence suggests these closely related receptors regulate different physiological outputs in the same cell, although...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.109.062018

    authors: McCoy KL,Traynelis SF,Hepler JR

    更新日期:2010-06-01 00:00:00

  • N-methyl-D-aspartate and brain-derived neurotrophic factor induce distinct profiles of extracellular signal-regulated kinase, mitogen- and stress-activated kinase, and ribosomal s6 kinase phosphorylation in cortical neurons.

    abstract::Stimulation of N-methyl-D-aspartate (NMDA) receptors is believed to underlie long-term memory formation, and excessive NMDA receptor activation has been linked to several neuropathological conditions. Phosphorylation and activation of p42/44 mitogen-activated protein kinase (ERK) is believed to mediate many of these e...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.104.005447

    authors: Rakhit S,Clark CJ,O'shaughnessy CT,Morris BJ

    更新日期:2005-04-01 00:00:00

  • Selective and synergistic inhibition of human immunodeficiency virus type 1 reverse transcriptase by a non-nucleoside inhibitor, MKC-442.

    abstract::In the search for 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine derivatives, we have found 6-benzyl-1-(ethoxymethyl)-5-isopropyl-uracil (MKC-442) to be a highly potent and selective inhibitor of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT). The IC50 value of MKC-442 for HIV-1 RT was 8 nM....

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Yuasa S,Sadakata Y,Takashima H,Sekiya K,Inouye N,Ubasawa M,Baba M

    更新日期:1993-10-01 00:00:00

  • Fusion polypeptides that inhibit exocytosis: fusing aptamer and cell-penetrating peptide technologies and pharmacologies.

    abstract::Cell-penetrating peptides are amphipathic or cationic oligopeptides able to transport covalently attached cargoes across cell membranes. Peptide aptamers are polypeptide fragments of endogenous proteins that mimic and thus perturb interactions with other cellular proteins. Combining aptamer and CPP technology can gene...

    journal_title:Molecular pharmacology

    pub_type: 评论,杂志文章

    doi:10.1124/mol.105.011429

    authors: Eiden LE

    更新日期:2005-04-01 00:00:00

  • In vitro binding characteristics of a new selective group II metabotropic glutamate receptor radioligand, [3H]LY354740, in rat brain.

    abstract::The in vitro binding of [3H]LY354740, the first high affinity group II-selective metabotropic glutamate (mGlu) receptor radioligand, was characterized in rat cortical, hippocampal, and thalamic membranes as well as in rat brain sections. [3H]LY354740 binding was saturable in all regions investigated. Nonspecific bindi...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Schaffhauser H,Richards JG,Cartmell J,Chaboz S,Kemp JA,Klingelschmidt A,Messer J,Stadler H,Woltering T,Mutel V

    更新日期:1998-02-01 00:00:00

  • New sigma-like receptor recognized by novel phenylaminotetralins: ligand binding and functional studies.

    abstract::Several novel phenylaminotetralins (PATs) cause functional changes in brain that are associated with binding to saturable, high affinity sites that are not identical to any known central nervous system receptor. These PATs were tested for their ability to cause receptor-mediated functional effects on tyrosine hydroxyl...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Booth RG,Wyrick SD,Baldessarini RJ,Kula NS,Myers AM,Mailman RB

    更新日期:1993-12-01 00:00:00

  • Trichosporin-B-III, an alpha-aminoisobutyric acid-containing peptide, causes Ca(2+)-dependent catecholamine secretion from adrenal medullary chromaffin cells.

    abstract::We examined the effect of trichosporin-B-III, an alpha-aminoisobutyric acid-containing antibiotic peptide consisting of 19 amino acid residues and a phenylalaninol, on catecholamine secretion from cultured bovine adrenal chromaffin cells. Incubation of the cells with trichosporin-B-III (3-20 microM) caused an increase...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Tachikawa E,Takahashi S,Furumachi K,Kashimoto T,Iida A,Nagaoka Y,Fujita T,Takaishi Y

    更新日期:1991-11-01 00:00:00

  • Expression of the pore-forming P2Z purinoreceptor in Xenopus oocytes injected with poly(A)+ RNA from murine macrophages.

    abstract::Extracellular ATP activates two distinct types of P2 purinoreceptor-mediated signaling pathways in macrophages, 1) the rapid formation of nonselective pores/channels in the plasma membrane and 2) a guanine nucleotide-binding protein-dependent stimulation of phosphotidylinositol-specific phospholipase C, with subsequen...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Nuttle LC,el-Moatassim C,Dubyak GR

    更新日期:1993-07-01 00:00:00

  • A novel constitutive androstane receptor-mediated and CYP3A-independent pathway of bile acid detoxification.

    abstract::Cytosolic sulfotransferase (SULT)-mediated sulfation plays an essential role in the detoxification of bile acids and is necessary to avoid pathological conditions, such as cholestasis, liver damage, and colon cancer. In this study, using transgenic mice bearing conditional expression of the activated constitutive andr...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.65.2.292

    authors: Saini SP,Sonoda J,Xu L,Toma D,Uppal H,Mu Y,Ren S,Moore DD,Evans RM,Xie W

    更新日期:2004-02-01 00:00:00

  • Identification of receptor domains that modify ligand binding to 5-hydroxytryptamine2 and 5-hydroxytryptamine1c serotonin receptors.

    abstract::Serotonin [5-hydroxytryptamine (5-HT)] receptors are distinguished pharmacologically by their characteristic affinities for agonists and antagonists. Two serotonin receptors, the 5-HT2 and 5-HT1c, share a number of pharmacologic and structural properties while differing in their affinities for certain agonists and ant...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Choudhary MS,Craigo S,Roth BL

    更新日期:1992-10-01 00:00:00

  • Real-time visualization of a fluorescent G(alpha)(s): dissociation of the activated G protein from plasma membrane.

    abstract::To study behavior of activated G(alpha)(s) in living cells, green fluorescent protein (GFP) was inserted within the internal amino acid sequence of G(alpha)(s) to generate a G(alpha)(s)-GFP fusion protein. The fusion protein maintained a bright green fluorescence and was identified by immunoblotting with antibodies ag...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.61.2.352

    authors: Yu JZ,Rasenick MM

    更新日期:2002-02-01 00:00:00

  • Long-term channel block is required to inhibit cellular transformation by human ether-à-go-go-related gene (hERG1) potassium channels.

    abstract::Both human ether-à-go-go-related gene (hERG1) and the closely related human ether-à-go-go (hEAG1) channel are aberrantly expressed in a large proportion of human cancers. In the present study, we demonstrate that transfection of hERG1 into mouse fibroblasts is sufficient to induce many features characteristic of malig...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.113.091439

    authors: Pier DM,Shehatou GS,Giblett S,Pullar CE,Trezise DJ,Pritchard CA,Challiss RA,Mitcheson JS

    更新日期:2014-08-01 00:00:00

  • Tricyclic antidepressants and dextromethorphan bind with higher affinity to the phencyclidine receptor in the absence of magnesium and L-glutamate.

    abstract::Recent studies from our laboratory have provided evidence that multiple states of the phencyclidine (PCP) receptor exist. In addition, several compounds such as PCP and the novel anticonvulsant MK-801 were found to inhibit binding more potently in the presence of Mg2+ and L-glutamate (L-GLU) than when these agents wer...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Sills MA,Loo PS

    更新日期:1989-07-01 00:00:00

  • The Pharmacology and Function of Receptors for Short-Chain Fatty Acids.

    abstract::Despite some blockbuster G protein-coupled receptor (GPCR) drugs, only a small fraction (∼ 15%) of the more than 390 nonodorant GPCRs have been successfully targeted by the pharmaceutical industry. One way that this issue might be addressed is via translation of recent deorphanization programs that have opened the pro...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章,评审

    doi:10.1124/mol.115.102301

    authors: Bolognini D,Tobin AB,Milligan G,Moss CE

    更新日期:2016-03-01 00:00:00

  • Functional coupling of human L-type Ca2+ channels and angiotensin AT1A receptors coexpressed in xenopus laevis oocytes: involvement of the carboxyl-terminal Ca2+ sensors.

    abstract::A human recombinant L-type Ca2+ channel (alpha1C,77) was coexpressed with the rat angiotensin AT1A receptor in Xenopus laevis oocytes. In oocytes expressing only alpha1C,77 channels, application of human angiotensin II (1-10 microM) did not affect the amplitude or kinetics of Ba2+ currents (IBa). In sharp contrast, in...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.54.6.1106

    authors: Oz M,Melia MT,Soldatov NM,Abernethy DR,Morad M

    更新日期:1998-12-01 00:00:00

  • Two alpha1-adrenergic receptor subtypes regulating the vasopressor response have differential roles in blood pressure regulation.

    abstract::To study the functional role of individual alpha1-adrenergic (AR) subtypes in blood pressure (BP) regulation, we used mice lacking the alpha1B-AR and/or alpha1D-AR with the same genetic background and further studied their hemodynamic and vasoconstrictive responses. Both the alpha1D-AR knockout and alpha1B-/alpha1D-AR...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.104.007500

    authors: Hosoda C,Koshimizu TA,Tanoue A,Nasa Y,Oikawa R,Tomabechi T,Fukuda S,Shinoura H,Oshikawa S,Takeo S,Kitamura T,Cotecchia S,Tsujimoto G

    更新日期:2005-03-01 00:00:00

  • Relative substrate activities of structurally related pteridine, quinazoline, and pyrimidine analogs for mouse liver folylpolyglutamate synthetase.

    abstract::Several structurally related series of folate analogs were studied as substrates for mouse liver folylpolyglutamate synthetase (FPGS). A comparison of the kinetics of the interaction of this enzyme with folate analogs that contained the quinazoline ring in place of the pteridine ring with those of the analogous pterid...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Moran RG,Colman PD,Jones TR

    更新日期:1989-11-01 00:00:00

  • Antimalarial activity of selected aromatic chelators. IV. Cation uptake by Plasmodium falciparum in the presence of oxines and siderochromes.

    abstract::The growth of Plasmodium falciparum, a human malaria parasite, is sensitive to inhibition by chelators of several types. The alkylthiocarbamates and 8-hydroxyquinoline at pharmacologic doses selectively inhibit glycolysis within 6 hr in parasitized erythrocytes. The mechanism attributed to these agents is through the ...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Scheibel LW,Stanton GG

    更新日期:1986-10-01 00:00:00

  • Mpl ligand increases P2Y1 receptor gene expression in megakaryocytes with no concomitant change in platelet response to ADP.

    abstract::The P2Y(1) receptor is responsible for the initiation of platelet aggregation in response to ADP and plays a key role in thrombosis. Although this receptor is expressed early in the platelet lineage, the regulation of its expression during megakaryocyte differentiation is unknown. In the mouse megakaryocytic cell line...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.60.5.1112

    authors: Hechler B,Toselli P,Ravanat C,Gachet C,Ravid K

    更新日期:2001-11-01 00:00:00

  • Block of high-threshold calcium channels by the synthetic polyamines sFTX-3.3 and FTX-3.3.

    abstract::A polyamine component of Agelenopsis aperta spider venom designated FTX is reported to be a selective antagonist of P-type calcium channels in the mammalian brain. Consequently, this component has frequently been used as a pharmacological tool to determine the presence, distribution, and function of P-type channels in...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Norris TM,Moya E,Blagbrough IS,Adams ME

    更新日期:1996-10-01 00:00:00

  • Direct and differential interaction of beta-arrestins with the intracellular domains of different opioid receptors.

    abstract::beta-arrestins have been shown to play important roles in regulation of signaling and desensitization of opioid receptors in many in vivo studies. The current study was carried out to measure the direct interaction of beta-arrestins with two functional intracellular domains, the third intracellular loop (I3L) and the ...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.59.4.758

    authors: Cen B,Xiong Y,Ma L,Pei G

    更新日期:2001-04-01 00:00:00

  • κ-Opioid receptor inhibition of calcium oscillations in spinal cord neurons.

    abstract::Mouse embryonic spinal cord neurons in culture exhibit spontaneous calcium oscillations from day in vitro (DIV) 6 through DIV 10. Such spontaneous activity in developing spinal cord contributes to maturation of synapses and development of pattern-generating circuits. Here we demonstrate that these calcium oscillations...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.111.071456

    authors: Kelamangalath L,Dravid SM,George J,Aldrich JV,Murray TF

    更新日期:2011-06-01 00:00:00

  • A new 2-pyrone derivative, 5-bromo-3-(3-hydroxyprop-1-ynyl)-2H-pyran-2-one, suppresses stemness in glioma stem-like cells.

    abstract::Glioma cells with stem cell properties, termed glioma stem-like cells (GSCs), have been linked to tumor formation, maintenance, and progression and are responsible for the failure of chemotherapy and radiotherapy. Because conventional glioma treatments often fail to eliminate GSCs completely, residual surviving GSCs a...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.112.078402

    authors: Kim RK,Kim MJ,Yoon CH,Lim EJ,Yoo KC,Lee GH,Kim YH,Kim H,Jin YB,Lee YJ,Cho CG,Oh YS,Gye MC,Suh Y,Lee SJ

    更新日期:2012-09-01 00:00:00

  • Coexpression with the inward rectifier K(+) channel Kir6.1 increases the affinity of the vascular sulfonylurea receptor SUR2B for glibenclamide.

    abstract::ATP-sensitive K(+) channels are closed by the hypoglycemic sulfonylureas like glibenclamide (GBC) and activated by a class of vasorelaxant compounds, the K(+) channel openers. These channels are octamers of Kir6.x and sulfonylurea receptor (SUR) subunits with 4:4 stoichiometry. The properties of the opener-sensitive K...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Russ U,Hambrock A,Artunc F,Löffler-Walz C,Horio Y,Kurachi Y,Quast U

    更新日期:1999-11-01 00:00:00

  • High affinity agonist binding to the dopamine D3 receptor: chimeric receptors delineate a role for intracellular domains.

    abstract::The dopamine D3 receptor, although structurally similar to the dopamine D2 receptor, has 100-fold higher affinity for agonists such as dopamine and quinpirole, when these receptors are expressed in 293 cells. Additionally, the D3 receptor has generally lower affinity for several antagonists than does the D2 receptor. ...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Robinson SW,Jarvie KR,Caron MG

    更新日期:1994-08-01 00:00:00

  • Tubulin-Binding 3,5-Bis(styryl)pyrazoles as Lead Compounds for the Treatment of Castration-Resistant Prostate Cancer.

    abstract::The microtubule-binding taxanes, docetaxel and cabazitaxel, are administered intravenously for the treatment of castration-resistant prostate cancer (CRPC) as the oral administration of these drugs is largely hampered by their low and highly variable bioavailabilities. Using a simple, rapid, and environmentally friend...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.119.118539

    authors: Liao VWY,Kumari A,Narlawar R,Vignarajan S,Hibbs DE,Panda D,Groundwater PW

    更新日期:2020-06-01 00:00:00

  • Identification of Abundant and Evolutionarily Conserved Opioid Receptor Circular RNAs in the Nervous System Modulated by Morphine.

    abstract::Circular RNAs (circRNAs) are a distinct category of single-stranded, covalently closed RNAs formed by backsplicing. The functions of circRNAs are incompletely known and are under active investigation. Here, we report that in addition to traditional linear mRNAs (linRNA), mouse, rat, and human opioid receptor genes gen...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:10.1124/mol.118.113977

    authors: Irie T,Shum R,Deni I,Hunkele A,Le Rouzic V,Xu J,Wilson R,Fischer GW,Pasternak GW,Pan YX

    更新日期:2019-08-01 00:00:00

  • In GH3 pituitary cells, acetylcholine and vasoactive intestinal peptide antagonistically modulate adenylate cyclase, cyclic AMP content, and prolactin secretion.

    abstract::In GH3 pituitary cell homogenates, acetylcholine (ACh) (IC50 200 nM) inhibits adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] activity in a concentration- and GTP-dependent manner. Maximal inhibition was obtained with 10 microM ACh and corresponded to approximately a 50% decrease in basal enzyme ac...

    journal_title:Molecular pharmacology

    pub_type: 杂志文章

    doi:

    authors: Onali P,Eva C,Olianas MC,Schwartz JP,Costa E

    更新日期:1983-09-01 00:00:00