Imperial College London
Portrait Picture

ProfessorAylinHanyaloglu

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Professor in Molecular Medicine
 
 
 
//

Contact

 

+44 (0)20 7594 2128a.hanyaloglu Website

 
 
//

Assistant

 

Miss Kiran Dosanjh +44 (0)20 7594 2176

 
//

Location

 

2009Institute of Reproductive and Developmental BiologyHammersmith Campus

//

Summary

 

Publications

Citation

BibTex format

@article{Baidya:2020:10.1074/jbc.RA120.013470,
author = {Baidya, M and Kumari, P and Dwivedi-Agnihotri, H and Pandey, S and Sokrat, B and Sposini, S and Chaturvedi, M and Srivastava, A and Roy, D and Hanyaloglu, AC and Bouvier, M and Shukla, AK},
doi = {10.1074/jbc.RA120.013470},
journal = {Journal of Biological Chemistry},
pages = {10153--10167},
title = {Genetically encoded intrabody sensors report the interaction and trafficking of β-arrestin 1 upon activation of G protein-coupled receptors},
url = {http://dx.doi.org/10.1074/jbc.RA120.013470},
volume = {295},
year = {2020}
}
Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Agonist stimulation of G protein-coupled receptors (GPCRs) typically leads to phosphorylation of GPCRs and binding to multifunctional proteins called β-arrestins (βarrs). The GPCR-βarr interaction critically contributes to GPCR desensitization, endocytosis, and downstream signaling, and GPCR-βarr complex formation can be used as a generic readout of GPCR and βarr activation. Although several methods are currently available to monitor GPCR-βarr interactions, additional sensors to visualize them may expand the toolbox and complement existing methods. We have previously described antibody fragments (FABs) that recognize activated βarr1 upon its interaction with the vasopressin V2 receptor C-terminal phosphopeptide (V2Rpp). Here, we demonstrate that these FABs efficiently report the formation of a GPCR-βarr1 complex for a broad set of chimeric GPCRs harboring the V2R C terminus. We adapted these FABs to an intrabody format by converting them to single-chain variable fragments (ScFvs) and used them to monitor the localization and trafficking of βarr1 in live cells. We observed that upon agonist simulation of cells expressing chimeric GPCRs, these intrabodies first translocate to the cell surface, followed by trafficking into intracellular vesicles. The translocation pattern of intrabodies mirrored that of βarr1, and the intrabodies co-localized with βarr1 at the cell surface and in intracellular vesicles. Interestingly, we discovered that intrabody sensors can also report βarr1 recruitment and trafficking for several unmodified GPCRs. Our characterization of intrabody sensors for βarr1 recruitment and trafficking expands currently available approaches to visualize GPCR-βarr1 binding, which may help decipher additional aspects of GPCR signaling and regulation.
AU  - Baidya,M
AU  - Kumari,P
AU  - Dwivedi-Agnihotri,H
AU  - Pandey,S
AU  - Sokrat,B
AU  - Sposini,S
AU  - Chaturvedi,M
AU  - Srivastava,A
AU  - Roy,D
AU  - Hanyaloglu,AC
AU  - Bouvier,M
AU  - Shukla,AK
DO  - 10.1074/jbc.RA120.013470
EP  - 10167
PY  - 2020///
SN  - 0021-9258
SP  - 10153
TI  - Genetically encoded intrabody sensors report the interaction and trafficking of β-arrestin 1 upon activation of G protein-coupled receptors
T2  - Journal of Biological Chemistry
UR  - http://dx.doi.org/10.1074/jbc.RA120.013470
UR  - https://www.ncbi.nlm.nih.gov/pubmed/32439801
UR  - http://hdl.handle.net/10044/1/80644
VL  - 295
ER  -
Download