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Phosphorylation of intracellular serine and threonine residues is the most important post translational modification of G protein-coupled receptors (GPCRs) also called heptahelical or seven transmembrane receptors (7TMR). After agonist exposure, these receptors acquire an active conformation, which is recognized by a family of highly specialized GPCR kinases (GRKs). Agonist-driven phosphorylation by GRKs regulates acute receptor desensitization, arrestin recruitment, internalization, post-activation signaling, long-term tolerance and drug addiction. Phosphosite-specific 7TM antibodies are designed to specifically detect agonist-activated GPCRs. In fact, recent work shows that ligand profiling using phosphosite-specific 7TM antibodies provides valuble information on ligand bias beyond that obtained with conventional ß-arrestin recruitment assays. Phosphosite-specific 7TM antibodies are novel tools for GPCR research that can be used to:

  • profile agonist properties of novel GPCR ligands
  • decipher the phosphorylation barcode of GPCRs
  • determine the spatial and temporal dynamics of receptor phosphorylation
  • identify relevant kinases and phosphatases for GPCR phosphorylation and dephosphoryation

Lifecycle3


Schematic representation of the G protein-coupled receptor phosphorylation / dephosphorylation cycle. GRK, G protein-coupled receptor kinase; PKC, protein kinase C; cPP1, catalytic subunit of protein phosphatase 1; R*, activated GPCR; CCP, clathrin-coated pit. 

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pS348/pS350-ACKR2 (phospho-Atypical Chemokine Receptor 2 Antibody)
pS348/pS350-ACKR2 (phospho-Atypical Chemokine...
Serine348/Serine350 (S348/S350) is major phosphorylation site of the Atypical Chemokine Receptor 2 (ACKR2). The pS348/pS350-ACKR2 antibody detects phosphorylation in response to agonists. S348/S350 phosphorylation is likely to be...
$ 350.00
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Agonist-induced Threonine317/Serine318 phosphorylation of the Dopamine Receptor 2.
pT317/pS318-D2 (phospho-Dopamine Receptor 2...
Threonine317/serine318 (T317/S318) is a major phosphorylation site of the D2 dopamine receptor. The pT317/pS318-D2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T317/S318...
$ 350.00
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Validation of the Dopamine Receptor 2 in transfected HEK293 cells.
D2 (non-phospho), Dopamine Receptor 2 Antibody
The non-phospho-D2 receptor antibody is directed against the third intracellular loop of mouse, rat and human D2 dopamine receptor. It detects both the long and short form of D2. It can be used to detect total D2 receptors in Western...
$ 350.00
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Agonist-induced Threonine324/Serine327 phosphorylation of the Complement C5a Receptor 1.
pT324/pS327-C5a1 (phospho-Complement C5a...
Threonine324/serine327 (T324/S327) is a major phosphorylation site of the C5a1 receptor. The pT324/pS327-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T324/S327...
$ 350.00
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Agonist-induced Serine332/Serine334 phosphorylation of the Complement C5a Receptor 1.
pS332/pS334-C5a1 (phospho-Complement C5a...
Serine332/serine334 (S332/S334) is a major phosphorylation site of the C5a1 receptor. The pS332/pS334-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S332/S334...
$ 350.00
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Validation of the Complement C5a Receptor 1 in transfected HEK293 cells.
C5a1 (non-phospho), Complement C5a Receptor 1...
The non-phospho-C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human C5a1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The non-phospho-C5a1...
$ 350.00
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Validation of the Complement C5a Receptor 2 in transfected HEK293 cells.
C5a2 (non-phospho), Complement C5a Receptor 2...
The non-phospho-C5a2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human C5a2. It can be used to detect total C5a2 receptors in Western blots independent of phosphorylation. The non-phospho-C5a2...
$ 250.00
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Agonist-induced Serine326/Threonine327 phosphorylation of the Complement C5a Receptor 2.
pS326/pT327-C5a2 (phospho-Complement C5a...
Serine326/Threonine327 (S326/T327) is major phosphorylation site of the Complement Peptide Receptor 2 (C5a2). The pS326/pT327-C5a2 antibody detects phosphorylation in response to agonists. S326/S327 phosphorylation is likely to be...
$ 350.00
NEW
Validation of the Atypical Chemokine Receptor 2 in transfected HEK293 cells.
ACKR2 (non-phospho), Atypical Chemokine...
The non-phospho-ACKR2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human ACKR2. It can be used to detect total ACKR2 receptors in Western blots independent of phosphorylation. The...
$ 250.00
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Agonist-induced Threonine355 phosphorylation of the Chemokine Receptor 10
pT355-CCR10 (phospho-Chemokine Receptor 10...
Threonine355 (T355) is a major phosphorylation site of the CCR10 receptor. The pT355-CCR10 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T355 phosphorylation is a key regulator...
$ 350.00
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Agonist-induced Threonine336 phosphorylation of the Complement C5a Receptor 1
pT336-C5a1 (phospho-Complement C5a Receptor 1...
Threonine336 (T336) is a major phosphorylation site of the C5a1 receptor. The pT336-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T336 phosphorylation is a key...
$ 350.00
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pS338/pT339-C5a1 (phospho-Complement C5a Receptor 1 Antibody)
pS338/pT339-C5a1 (phospho-Complement C5a...
Serine338/Threonine339 (S338/T339) is a major phosphorylation site of the C5a1 receptor. The pS338/pT339-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S338/T339...
$ 350.00
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Agonist-induced Threonine342 phosphorylation of the Complement C5a Receptor 1
pT342-C5a1 (phospho-Complement C5a Receptor 1...
Threonine342 (T342) is a major phosphorylation site of the C5a1 receptor. The pT342-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T342 phosphorylation is a key...
$ 350.00
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Agonist-induced Threonine343/Serine347/Threonine348 phosphorylation of the 5-Hydroxytryptamine Receptor 4
pT343/pS347/pT348-5-HT4 (phospho-5-HT Receptor...
Threonine343/Serine347/Threonine348 is a major phosphorylation site of the 5-HT4 receptor. The pT343/pS347/pT348-5-HT4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T343/S347/T348...
$ 350.00
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Agonist-induced Serine354/Threonine355 phosphorylation of the 5-Hydroxytryptamine Receptor 4
pS354/pT355-5-HT4 (phospho-5-HT Receptor 4...
Serine354/Threonine355 is a major phosphorylation site of the 5-HT4 receptor. The pS354/pT355-5-HT4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S354/T355 phosphorylation is a key...
$ 350.00
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Agonist-induced Serine355/Serine356 phosphorylation of the β2 Adrenoceptor
pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)
Serine355/Serine356 (S355/S356) is a major phosphorylation site of the β2 adrenoceptor. The pS355/pS356-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S355/S356...
$ 350.00
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For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

For further reading refer to: Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636.... read more »
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For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

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