Imperial College London
Alternate

Professor Sir Steve Bloom FMedSci, FRS

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Departmental Academic REF2014 Lead
 
 
 
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Contact

 

+44 (0)20 7594 9048s.bloom Website

 
 
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Assistant

 

Ms Keda Price-Cousins +44 (0)20 7594 9048

 
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Location

 

6N3Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{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, S and Tan, T and Tomas, A},
doi = {10.1101/2021.05.09.443291},
journal = {Molecular Metabolism},
pages = {1--11},
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},
volume = {53},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - ObjectivesReceptor 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 (GCG) is important for carbohydrate and lipid metabolism.MethodsSubcellular localisation of GCGR in the presence and absence of RAMP2 was investigated using confocal microscopy, trafficking 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 and the trafficking inhibitor monensin were used to investigate the effect of a halt in recycling of internalised proteins on GCGR subcellular localisation and signalling in the absence of RAMP2. NanoBiT complementation and cyclic AMP assays were used to study the functional effect of RAMP2 on the 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.ResultsGCGR is predominantly localised at the plasma membrane in the absence of RAMP2 and exhibits remarkably slow internalisation in response to agonist stimulation. Rapid intracellular accumulation of GCG-stimulated GCGR in cells lacking WASH complex or in the presence of monensin 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 of RAMP2 confers a bias away from β-arrestin-2 recruitment coupled to increased activation of Gαs proteins at endosomes. This is associated with increased short-term efficacy for glucagon-stimulated
AU  - McGlone,ER
AU  - Manchanda,Y
AU  - Jones,B
AU  - Pickford,P
AU  - Inoue,A
AU  - Carling,D
AU  - Bloom,S
AU  - Tan,T
AU  - Tomas,A
DO  - 10.1101/2021.05.09.443291
EP  - 11
PY  - 2021///
SN  - 2212-8778
SP  - 1
TI  - Receptor Activity-Modifying Protein 2 (RAMP2) alters glucagon receptor trafficking in hepatocytes with functional effects on receptor signalling
T2  - Molecular Metabolism
UR  - http://dx.doi.org/10.1101/2021.05.09.443291
UR  - https://www.sciencedirect.com/science/article/pii/S2212877821001411?via%3Dihub
UR  - http://hdl.handle.net/10044/1/90405
VL  - 53
ER  -
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