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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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Validation of the Glucagon-Like Peptide 2 Receptor in transfected HEK293 cells
GLP2 (non-phospho) Glucagon-Like Peptide 2...
The non-phospho GLP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human GLP2 receptor. It can be used to detect total GLP2 receptors in Western blots independent of phosphorylation. It can also be...
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NEW
Immunohistochemical identification of Glucagon-Like Peptide 2 Receptor in enteric ganglion
GLP2 (IHC-grade), Glucagon-Like Peptide 2...
The non-phospho GLP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human GLP2 receptor. It can be used to detect total GLP2 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the Glucagon Receptor in transfected HEK293 cells
GCGR (non-phospho) Glucagon Receptor Antibody
The non-phospho GCGR antibody is directed against the distal end of the carboxyl-terminal tail of human GCGR. It can be used to detect total GCGR receptors in Western blots independent of phosphorylation. It can also be used to isolate...
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Validation of the Gastric Inhibitory Polypeptide Receptor in transfected HEK293 cells
GIPR (non-phospho) Gastric Inhibitory...
The non-phospho GIPR antibody is directed against the distal end of the carboxyl-terminal tail human GIPR. It can be used to detect total GIPR receptors in Western blots independent of phosphorylation. It can also be used to isolate and...
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Agonist-induced Serine363/Threonine364 phosphorylation of the Bombesin Receptor 2
pS363/pT364-BB2 (phospho-Bombesin Receptor 2...
Serine363/Threonine364 (S363/T364) is major phosphorylation site of the Bombesin Receptor 2 (BB2). The pS363/pT364-BB2 antibody detects phosphorylation in response to agonists. S363/T364 phosphorylation is likely to be involved in...
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Agonist-induced Threonine359/Serine360 phosphorylation of the Bombesin Receptor 2
pT359/pS360-BB2 (phospho-Bombesin Receptor 2...
Threonine359/Serine360 (T359/S360) is major phosphorylation site of the Bombesin Receptor 2 (BB2). The pT359/pS360-BB2 antibody detects phosphorylation in response to agonists. T359/S360 phosphorylation is likely to be involved in...
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Agonist-induced Threonine360/Serine361 phosphorylation of the Bombesin Receptor 3
pT360/pS361-BB3 (phospho-Bombesin Receptor 3...
Threonine360/Serine361 (T360/S361) is major phosphorylation site of the Bombesin Receptor 3 (BB3). The pT360/pS361-BB3 antibody detects phosphorylation in response to agonists. T360/S361 phosphorylation is likely to be involved in...
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Agonist-induced Threonine363/Threonine364 phosphorylation of the Bombesin Receptor 3
pT363/pT364-BB3 (phospho-Bombesin Receptor 3...
Threonine363/Threonine360 (T363/T364) is major phosphorylation site of the Bombesin Receptor 3 (BB3). The pT363/pT364-BB3 antibody detects phosphorylation in response to agonists. T363/T364 phosphorylation is likely to be involved in...
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Agonist-induced Threonine370 phosphorylation of the Bombesin Receptor 3
pT370-BB3 (phospho-Bombesin Receptor 3 Antibody)
Threonine370 (T370) is major phosphorylation site of the Bombesin Receptor 3 (BB3). The pT370-BB3 antibody detects phosphorylation in response to agonists. T370 phosphorylation is likely to be involved in efficient ligand sequestration...
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Validation of the Motilin Receptor in transfected HEK293 cells
MLNR (non-phospho) Motilin Receptor Antibody
The non-phospho MLNR antibody is directed against the distal end of the carboxyl-terminal tail human MLNR receptor. It can be used to detect total MLNR receptors in Western blots independent of phosphorylation. It can also be used to...
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Agonist-induced Serine298/Serine300 phosphorylation of the M4 Muscarinic Acetylcholine Receptor
pS298/pS300-M4 (phospho-M4 Muscarinic...
Serine298/Serine300 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS298/pS300-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S298/S300...
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Agonist-induced Serine492/Serine493 phosphorylation of the Parathyroid Hormone Receptor 1
pS492/pS493-PTH1 (phospho-Parathyroid Hormone...
Serine492/Serine493 (S492/S493) is major phosphorylation site of the Parathyroid Hormone Receptor 1 (PTH1). The pS492/pS493-PTH1 antibody detects phosphorylation in response to agonists. S492/S493 phosphorylation is likely to be involved...
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Agonist-induced Serine501/Threonine503/Serine504 phosphorylation of the Parathyroid Hormone Receptor 1
pS501/pT503/pS504-PTH1 (phospho-Parathyroid...
Serine501/Threonine503/Serine504 (S501/T503/S504) is major phosphorylation site of the Parathyroid Hormone Receptor 1 (PTH1). The pS501pT503/pS504-PTH1 antibody detects phosphorylation in response to agonists. S501T503/S504...
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pS331/pT332-LPA2 (phospho-Lysophosphatidic Acid Receptor 2 Antibody)
pS331/pT332-LPA2 (phospho-Lysophosphatidic Acid...
Serine331/Threonine332 (S331/T332) is major phosphorylation site of the Lysophosphatidic Acid Receptor 2 (LPA2). pS331/pT332-LPA2 antibody detects phosphorylation in response to agonists. S331/T332 phosphorylation is likely to be...
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CALCR (GP-non-phospho) Calcitonin Receptor-Like Receptor Antibody, Guinea Pig
CALCR (GP-non-phospho) Calcitonin Receptor-Like...
The non-phospho-CALCR receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CALCR. It can be used to detect total CALCR receptors in Western blots independent of phosphorylation. The CALCR antibody...
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Citations
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Immunohistochemical Identification of Agonist-induced Serine425 phosphorylation of the Cannabinoid Receptor 1 in mouse brain
pS425-CB1 (IHC-grade phospho-Cannabinoid...
Serine425 (S425) is major phosphorylation site of the Cannabinoid Receptor 1 (CB1). The pS425-CB1 antibody detects phosphorylation in response to agonists. S425 phosphorylation is likely to be involved in efficient ligand sequestration...
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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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