Internalized Receptor for Glucose-dependent Insulinotropic Peptide stimulates adenylyl cyclase on early endosomes
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Internalized Receptor for Glucose-dependent Insulinotropic Peptide stimulates adenylyl cyclase on early endosomes. / Ismail, Sadek; Gherardi, Marie-Julie; Froese, Alexander; Zanoun, Madjid; Gigoux, Véronique; Clerc, Pascal; Gaits-Iacovoni, Frederique; Steyaert, Jan; Nikolaev, Viacheslav O; Fourmy, Daniel.
In: BIOCHEM PHARMACOL, Vol. 120, 15.11.2016, p. 33-45.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Internalized Receptor for Glucose-dependent Insulinotropic Peptide stimulates adenylyl cyclase on early endosomes
AU - Ismail, Sadek
AU - Gherardi, Marie-Julie
AU - Froese, Alexander
AU - Zanoun, Madjid
AU - Gigoux, Véronique
AU - Clerc, Pascal
AU - Gaits-Iacovoni, Frederique
AU - Steyaert, Jan
AU - Nikolaev, Viacheslav O
AU - Fourmy, Daniel
N1 - Copyright © 2016 Elsevier Inc. All rights reserved.
PY - 2016/11/15
Y1 - 2016/11/15
N2 - Until very recently, G-protein dependent signal of GPCRs was thought to originate exclusively from the plasma membrane and internalized GPCRs were considered silent. Here, we demonstrated that, once internalized and located in the membrane of early endosomes, glucose-dependent Insulinotropic receptor (GIPR) continues to trigger production of cAMP and PKA activation. Direct evidence is based on identification of the active form of Gαs in early endosomes containing GIPR using a genetically encoded GFP tagged nanobody, and on detection of a distinct FRET signal accounting for cAMP production at the surface of endosomes containing GIP, compared to endosomes without GIP. Furthermore, decrease of the sustained phase of cAMP production and PKA activation kinetics as well as reversibility of cAMP production and PKA activity following GIP washout in cells treated with a pharmacological inhibitor of GIPR internalization, and continuous increase of cAMP level over time in the presence of dominant-negative Rab7, which causes accumulation of early endosomes in cells, were noticed. Hence the GIPR joins the few GPCRs which signal through G-proteins both at plasma membrane and on endosomes.
AB - Until very recently, G-protein dependent signal of GPCRs was thought to originate exclusively from the plasma membrane and internalized GPCRs were considered silent. Here, we demonstrated that, once internalized and located in the membrane of early endosomes, glucose-dependent Insulinotropic receptor (GIPR) continues to trigger production of cAMP and PKA activation. Direct evidence is based on identification of the active form of Gαs in early endosomes containing GIPR using a genetically encoded GFP tagged nanobody, and on detection of a distinct FRET signal accounting for cAMP production at the surface of endosomes containing GIP, compared to endosomes without GIP. Furthermore, decrease of the sustained phase of cAMP production and PKA activation kinetics as well as reversibility of cAMP production and PKA activity following GIP washout in cells treated with a pharmacological inhibitor of GIPR internalization, and continuous increase of cAMP level over time in the presence of dominant-negative Rab7, which causes accumulation of early endosomes in cells, were noticed. Hence the GIPR joins the few GPCRs which signal through G-proteins both at plasma membrane and on endosomes.
U2 - 10.1016/j.bcp.2016.09.009
DO - 10.1016/j.bcp.2016.09.009
M3 - SCORING: Journal article
C2 - 27641811
VL - 120
SP - 33
EP - 45
JO - BIOCHEM PHARMACOL
JF - BIOCHEM PHARMACOL
SN - 0006-2952
ER -
