Mutation of a conserved serine in TM4 of opioid receptors confers full agonistic properties to classical antagonists.

Claude, Patricia A.; Wotta, Diane R.; Zhang, X. H.; Prather, Paul L.; McGinn, Therese M.; Erickson, Leonard C.; Loh, Horace H.; Law, Ping Yee

doi:10.1073/pnas.93.12.5715

Cited by 58 publications

(50 citation statements)

References 26 publications

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“…When drugs were incubated with hCHO cells at this concentration for 20 min, dihydroetorphine, sufentanil, etorphine, etonitazine, and DAMGO caused increased receptor phosphorylation producing values 6 -8-fold above basal levels. Methadone, morphine, meperidine, DADL, ␤-endorphin , Met-enkephalin, Leu-enkephalin, and dynorphin A (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) produced intermediate effects, ϳ2-5 fold above basal levels. Neither the partial agonist buprenorphine, the largely agonists ketocyclazocine and ␣-neoendorphin, the mixed agonist/antagonist pentazocine, the mixed / agonist butorphanol, the weak agonists meperidine and 3-m-hydroxy-5-phenylmorphan, or the antagonists naloxaonazine and LY255582 had any robust effect on receptor phosphorylation.…”

Section: Resultsmentioning

confidence: 99%

“…However, mutagenesis studies and studies with receptor chimeras support the idea that different receptor features could be involved in recognition of these different ligand classes (4 -6). Mutations that change naloxone from a full antagonist to a partial agonist can leave the intrinsic activity of opioid peptides unchanged, for example see Claude et al (7). These differences raise the possibility that receptor occupancies by opioid drugs of different classes could alter the conformation of the receptor in distinct fashions.…”

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confidence: 99%

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μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

Zhang

Yin

et al. 1997

Journal of Biological Chemistry

201

142

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Opioid receptors are G-protein coupled receptors that mediate the potent analgesic actions and addictive properties of morphine-derived compounds. Under physiological conditions, these receptors interact with endogenous opioid peptides to modulate pain-controlling pathways and circuits that modulate behaviors including mood and reward (2). opioid receptors interact with rapidly acting opioid drugs, such as heroin, to produce marked euphoria and behavioral reward. They are also primary targets of the slower and longer acting opioids, such as methadone and LAAM, 1 that represent the best current substitution therapeutics for opiate addiction (3).receptors desensitize after repeated stimulation by opioid agonists, in fashions that display similarities to desensitizing events noted for other G-protein coupled receptors. Agonistinduced receptor desensitization can be correlated with receptor phosphorylation. receptors display naloxone-reversible phosphorylation and desensitize after morphine or DAMGO treatments (1). Both of these agonist-induced events are insensitive to pretreatments with the protein kinase C inhibitor staurosporine, which inhibits phorbol ester-induced receptor phosphorylation and desensitization.Many opiates and opioid ligands can recognize receptors with high affinities. Plant-derived alkaloids, synthetic compounds of several classes, and endogenous opioid peptides can function as agonists, partial agonists, or antagonists with a range of abilities to induce or block induction of analgesia and euphoria. However, mutagenesis studies and studies with receptor chimeras support the idea that different receptor features could be involved in recognition of these different ligand classes (4 -6). Mutations that change naloxone from a full antagonist to a partial agonist can leave the intrinsic activity of opioid peptides unchanged, for example see Claude et al. (7). These differences raise the possibility that receptor occupancies by opioid drugs of different classes could alter the conformation of the receptor in distinct fashions. Some of these differences could render the receptor an improved or a worse substrate for kinases and phosphatases and thus directly confer different susceptibilities to phosphorylation or dephosphorylation. Alternatively, selective activation of different G-protein classes by different agonists could trigger different receptor phosphorylation and desensitization events.The current study thus investigates the effects of opioid ligands of various classes and varying intrinsic activities on receptor activation, phosphorylation, and desensitization using human receptors expressed in CHO cells (hCHO), and the receptors coexpressed in Xenopus oocytes with a G-protein linked K ϩ channel. The results support striking parallels between opioid efficacies in opening ion channels and inhibiting

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

Zhang

Yin

et al. 1997

Journal of Biological Chemistry

201

142

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“…Strader et al (1989) explained their ®ndings by suggesting the existence of overlapping binding sites for agonists and antagonists. Claude et al (1996) hypothesize that antagonists with high structural homology to agonists are capable of agonist activity if the receptor is permissive. This was based on their observation that the mutation of conserved serine residues (Ser 177 Leu or Ser 196 Leu) in transmembrane domain IV of m-opioid receptors confers full agonistic properties to peptide (TIPPj) and alkaloid antagonists (naloxone, naltrexone and naltriben).…”

Section: Partial Agonist Activity Of Antagonists At Mutant G-protein-mentioning

confidence: 99%

G‐protein activation by putative antagonists at mutant Thr³⁷³Lys α_2A adrenergic receptors

Wurch

Colpaert

Pauwels

1999

British J Pharmacology

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1 Replacement of a threonine by a lysine at position 373 in the C-terminal portion of the third intracellular loop of the human a 2A -adrenergic receptor (a 2A AR) has been reported to generate a constitutively active mutant receptor in analogy with similar mutations in the a 1B and b 2 AR (Ren et al., 1993). In the present study, the mutant Thr 373 Lys a 2A AR receptor was investigated by measuring the formation of inositol phosphates in either the absence or presence of mouse G a15 protein in Cos-7 cells. 2 Increased a nity, potency and/or e cacy for the agonists [(7)-adrenaline, UK 14304, clonidine, guanabenz and oxymetazoline] was observed, consistent with a precoupled mutant a 2A AR: Gprotein state. The basal inositol phosphates response was similar at the wild-type (wt) and mutant a 2A AR, but was enhanced at the mutant a 2A AR upon co-expression with the mouse G a15 protein.This enhanced response could not be attenuated in the presence of any of the tested a 2 AR antagonists (10 mM), suggesting that inverse agonist activity did not occur at the mutant a 2A AR. 4 The partial agonist e ect of SKF 86466 was resistant to pertussis toxin treatment (100 ng ml 71 ) and not a ected by co-expression of the rat G ai1 protein. It was virtually absent in the presence of 10 mM RS 15385. SKF 86466 was without intrinsic activity upon co-expression of the mouse G aq protein.5 Some putative a 2 AR antagonists exerted a partial agonist activity that was highly dependent on the presence of speci®c G-protein a-subunits, suggesting that these ligands cause selective G-protein activation at the mutant a 2A AR.

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“…In the ␤-adrenergic (40) and 5HT2A (41) Mutations in a number of GPCRs have been described that confer agonism to antagonists. Mutations in TM3 of the AT1 angiotensin receptor (39,42), rhodopsin (43), and the ␤-adrenergic receptor (40), in TM4 of -and ␦-opioid receptors (44), in TM6 of the -opioid receptor (45), in IC3 of the ␣ 2A -adrenergic receptor (46), and in IC2 of the V2 vasopressin receptor (47) conferred agonistic activity to antagonist ligands. The present study is the first description of a mutation in an extracellular domain that confers agonism to an antagonist.…”

Section: Chicken Gnrh Receptor Ec2 Confers Agonism To Antagonistsmentioning

confidence: 99%

A Chicken Gonadotropin-releasing Hormone Receptor That Confers Agonist Activity to Mammalian Antagonists

Sun

Flanagan

Illing

et al. 2001

Journal of Biological Chemistry

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Mammalian receptors for gonadotropin-releasing hormone (GnRH) have over 85% sequence homology and similar ligand selectivity. Biological studies indicated that the chicken GnRH receptor has a distinct pharmacology, and certain antagonists of mammalian GnRH receptors function as agonists. To explore the structural determinants of this, we have cloned a chicken pituitary GnRH receptor and demonstrated that it has marked differences in primary amino acid sequence (59% homology) and in its interactions with GnRH analogs. The chicken GnRH receptor had high affinity for mammalian GnRH (K i 4.1 ؎ 1.2 nM) , similar to the human receptor (K i 4.8 ؎ 1.2 nM). But, in contrast to the human receptor, it also had high affinity for chicken GnRH or D-isopropyl-Lys 6 moieties, functioned as pure antagonists in the human receptor but were full or partial agonists in the chicken receptor. This suggests that the Lys side chain interacts with functional groups of the chicken GnRH receptor to stabilize it in the active conformation and that these groups are not available in the activated human GnRH receptor. Substitution of the human receptor extracellular loop two with the chicken extracellular loop two identified this domain as capable of conferring agonist activity to mammalian antagonists. Although functioning of antagonists as agonists has been shown to be species-dependent for several GPCRs, the dependence of this on an extracellular domain has not been described.

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Mutation of a conserved serine in TM4 of opioid receptors confers full agonistic properties to classical antagonists.

Cited by 58 publications

References 26 publications

μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

G‐protein activation by putative antagonists at mutant Thr³⁷³Lys α_2A adrenergic receptors

A Chicken Gonadotropin-releasing Hormone Receptor That Confers Agonist Activity to Mammalian Antagonists

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Mutation of a conserved serine in TM4 of opioid receptors confers full agonistic properties to classical antagonists.

Cited by 58 publications

References 26 publications

μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

μ Opioid Receptor Phosphorylation, Desensitization, and Ligand Efficacy

G‐protein activation by putative antagonists at mutant Thr373Lys α2A adrenergic receptors

A Chicken Gonadotropin-releasing Hormone Receptor That Confers Agonist Activity to Mammalian Antagonists

Contact Info

Product

Resources

About

G‐protein activation by putative antagonists at mutant Thr³⁷³Lys α_2A adrenergic receptors