Characterization of Glucagon-Like Peptide-1 Receptor β-Arrestin 2 Interaction: A High-Affinity Receptor Phenotype

Jørgensen, Rasmus; Martini, Lene; Schwartz, Thue W.; Elling, Christian

doi:10.1210/me.2004-0312

Cited by 68 publications

(74 citation statements)

References 43 publications

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“…Moreover, the fraction of receptor molecules in the high agonist affinity state in the presence of both G protein and arrestin was found to be directly proportional to the intrinsic activity of the agonist (Samama et al, 1993;Simons et al, 2004). Similar observations with the N-formyl peptide receptor Key et al, 2001Key et al, , 2003 and glucagon-like peptide-1 receptor (Jorgensen et al, 2005) strongly indicate that this is a general rule for the GPCR family. The translocation of arrestin-GFP fusion proteins to the plasma membrane in response to receptor activation is a popular indicator of arrestin binding to the receptor in living cells .…”

Section: Do G Proteins and Arrestins Require The Same "Active" Receptsupporting

confidence: 70%

The structural basis of arrestin-mediated regulation of G-protein-coupled receptors

Gurevich

2006

Pharmacology & Therapeutics

410

446

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The 4 mammalian arrestins serve as almost universal regulators of the largest known family of signaling proteins, G-protein-coupled receptors (GPCRs). Arrestins terminate receptor interactions with G proteins, redirect the signaling to a variety of alternative pathways, and orchestrate receptor internalization and subsequent intracellular trafficking. The elucidation of the structural basis and fine molecular mechanisms of the arrestin-receptor interaction paved the way to the targeted manipulation of this interaction from both sides to produce very stable or extremely transient complexes that helped to understand the regulation of many biologically important processes initiated by active GPCRs. The elucidation of the structural basis of arrestin interactions with numerous nonreceptor-binding partners is long overdue. It will allow the construction of fully functional arrestins in which the ability to interact with individual partners is specifically disrupted or enhanced by targeted mutagenesis. These "custom-designed" arrestin mutants will be valuable tools in defining the role of various interactions in the intricate interplay of multiple signaling pathways in the living cell. The identification of arrestin-binding sites for various signaling molecules will also set the stage for designing molecular tools for therapeutic intervention that may prove useful in numerous disorders associated with congenital or acquired disregulation of GPCR signaling.

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Section: Do G Proteins and Arrestins Require The Same "Active" Receptsupporting

confidence: 70%

The structural basis of arrestin-mediated regulation of G-protein-coupled receptors

Gurevich

2006

Pharmacology & Therapeutics

410

446

View full text Add to dashboard Cite

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“…By stabilizing a receptor's active conformation, arrestin increases its affinity for agonist (Gurevich et al, 1997;Key et al, 2001;Martini et al, 2002;Key et al, 2003;Jorgensen et al, 2005). As shown in Table 1, arrestin caused a profound increase in agonist affinity for all of the TRH receptors studied.…”

Section: Discussionmentioning

confidence: 89%

Arrestin Binds to Different Phosphorylated Regions of the Thyrotropin-Releasing Hormone Receptor with Distinct Functional Consequences

Jones

Hinkle²

2008

Mol Pharmacol

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Arrestin binding to agonist-occupied phosphorylated G protein-coupled receptors typically increases the affinity of agonist binding, increases resistance of receptor-bound agonist to removal with high acid/salt buffer, and leads to receptor desensitization and internalization. We tested whether thyrotropinreleasing hormone (TRH) receptors lacking phosphosites in the C-terminal tail could form stable and functional complexes with arrestin. Fibroblasts from mice lacking arrestins 2 and 3 were used to distinguish between arrestin-dependent and -independent effects. Arrestin did not promote internalization or desensitization of a receptor that had key Ser/Thr phosphosites mutated to Ala (4Ala receptor). Nevertheless, arrestin greatly increased acid/salt resistance and the affinity of 4Ala receptor for TRH. Truncation of 4Ala receptor just distal to the key phosphosites (4AlaStop receptor) abolished arrestin-dependent acid/salt resistance but not the effect of arrestin on agonist affinity. Arrestin formed stable complexes with activated wild-type and 4Ala receptors but not with 4AlaStop receptor, as measured by translocation of arrestin-green fluorescent protein to the plasma membrane or chemical cross-linking. An arrestin mutant that does not interact with clathrin and AP2 did not internalize receptor but still promoted high affinity TRH binding, acid/salt resistance, and desensitization. A sterically restricted arrestin mutant did not cause receptor internalization or desensitization but did promote acid/salt resistance and high agonist affinity. The results demonstrate that arrestin binds to proximal or distal phosphosites in the receptor tail. Arrestin binding at either site causes increased agonist affinity and acid/salt resistance, but only the proximal phosphosites evoke the necessary conformational changes in arrestin for receptor desensitization and internalization.

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“…Because β-arrestin recruitment is dependent on receptor phosphorylation by GRKs, β2-AR/Rluc was cotransfected with GRK 2 or GRK 5 for potential optimization of receptor phosphorylation. Jorgensen et al 20 have shown that this enhances the sensitivity of the BRET 2 assay. Moreover, McGraw and Liggett 21 demonstrated that a relatively low level of GRK 2 expression in human airway smooth muscle (HASM) is associated with an attenuated rate of short-term agonist-promoted β2-AR desensitization.…”

Section: Resultsmentioning

confidence: 98%

Bioluminescence Resonance Energy Transfer as a Screening Assay: Focus on Partial and Inverse Agonism

Elster¹,

Elling²,

Heding³

2007

SLAS Discovery

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The reported data for compound screening with the bioluminescence resonance energy transfer (BRET 2 ) assay is very limited, and several questions remain unaddressed, such as the behavior of agonists. Eleven β2 adrenergic receptor (β2-AR) agonists were tested for full or partial agonism in an improved version of the receptor/β-arrestin2 BRET 2 assay and in 2 cyclic adenosine monophosphate (cAMP) assays (column cAMP assay and ALPHAscreen™ cAMP assay). Tested in the highly sensitive ALPHAscreen™ cAMP assay, all selected agonists behaved as full agonists, using isoproterenol as a reference compound. In the less sensitive column cAMP assay, ephedrine and dopamine had a clear partial response. For the BRET 2 assay, a highly graded picture was obtained. Moreover, β2-AR antagonists were tested for inverse agonism. Pronounced inverse agonism was detected in the ALPHAscreen™ cAMP assay. Only marginal inverse agonistic responses were seen for alprenolol and pindolol in the column cAMP assay, and no inverse agonism was seen in the BRET 2 assay. For the β2-AR, the BRET 2 assay may be superior for secondary screening of agonists where a separation of full and partial agonists is needed and the ALPHAscreen™ cAMP assay may be preferred for primary screening of agonists where all receptor activating compounds are desired. (Journal of Biomolecular Screening 2007:41-49)

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Characterization of Glucagon-Like Peptide-1 Receptor β-Arrestin 2 Interaction: A High-Affinity Receptor Phenotype

Cited by 68 publications

References 43 publications

The structural basis of arrestin-mediated regulation of G-protein-coupled receptors

The structural basis of arrestin-mediated regulation of G-protein-coupled receptors

Arrestin Binds to Different Phosphorylated Regions of the Thyrotropin-Releasing Hormone Receptor with Distinct Functional Consequences

Bioluminescence Resonance Energy Transfer as a Screening Assay: Focus on Partial and Inverse Agonism

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