In this section
@unpublished{McGlone:2021:10.1101/2021.05.09.443291,
author = {McGlone, ER and Manchanda, Y and Jones, B and Pickford, P and Inoue, A and Carling, D and Bloom, SR and Tan, T and Tomas, A},
doi = {10.1101/2021.05.09.443291},
title = {Receptor Activity-Modifying Protein 2 (RAMP2) alters glucagon receptor trafficking in hepatocytes with functional effects on receptor signalling},
url = {http://dx.doi.org/10.1101/2021.05.09.443291},
year = {2021}
}
TY - UNPB
AB - <jats:title>Abstract</jats:title><jats:sec><jats:title>Objectives</jats:title><jats:p>Receptor Activity-Modifying Protein 2 (RAMP2) is a chaperone protein which allosterically binds to and interacts with the glucagon receptor (GCGR). The aims of this study were to investigate the effects of RAMP2 on GCGR trafficking and signalling in the liver, where glucagon is important for carbohydrate and lipid metabolism.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>Subcellular localisation of GCGR in the presence and absence of RAMP2 was investigated using confocal microscopy, trafficking assays and radioligand binding assays in human embryonic kidney (HEK293T) and human hepatoma (Huh7) cells. Mouse embryonic fibroblasts (MEFs) lacking Wiskott Aldrich Syndrome protein and scar homologue (WASH) complex were used to investigate the effect of a halt in recycling of internalised proteins on GCGR signalling in the absence of RAMP2. NanoBiT complementation and cyclic AMP assays were used to study the functional effect of RAMP2 on recruitment and activation of GCGR signalling mediators. Response to hepatic RAMP2 up-regulation in lean and obese adult mice using a bespoke adeno-associated viral vector was also studied.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>GCGR is predominantly localised at the plasma membrane in the absence of RAMP2 and exhibits remarkably slow internalisation in response to agonist stimulation. Rapid intracellular retention of glucagon-stimulated GCGR in cells lacking WASH complex indicates that activated GCGRs undergo continuous cycles of internalisation and recycling despite apparent GCGR plasma membrane localisation up to 40 minutes post-stimulation. Co-expression of RAMP2 induces GCGR internalisation both basally and in response to agonist-stimulation. The intracellular retention of GCGR in the presence
AU - McGlone,ER
AU - Manchanda,Y
AU - Jones,B
AU - Pickford,P
AU - Inoue,A
AU - Carling,D
AU - Bloom,SR
AU - Tan,T
AU - Tomas,A
DO - 10.1101/2021.05.09.443291
PY - 2021///
TI - Receptor Activity-Modifying Protein 2 (RAMP2) alters glucagon receptor trafficking in hepatocytes with functional effects on receptor signalling
UR - http://dx.doi.org/10.1101/2021.05.09.443291
UR - https://doi.org/10.1101/2021.05.09.443291
ER -