2026-04-17T06:13:25Zhttps://recercat.cat/oai/request
oai:recercat.cat:10230/556712025-12-20T16:50:31Zcom_2072_6col_2072_452952
RECERCAT author Baidya, Mithu author Chaturvedi, Madhu author Dwivedi, Hemlata author Ranjan, Ashutosh author Devost, Dominic author Namkung, Yoon author Stepniewski, Tomasz Maciej, 1988- author Pandey, Shubhi author Baruah, Minakshi author Panigrahi, Bhanupriya author Sarma, Parishmita author Yadav, Manish K. author Maharana, Jagannath author Banerjee, Ramanuj author Kawakami, Kouki author Inoue, Asuka author Selent, Jana author Laporte, Stéphane A. author Hébert, Terence E. author Shukla, Arun K. 2023-02-08T07:23:51Z2023-02-08T07:23:51Z2022 Agonist-induced phosphorylation of G protein-coupled receptors (GPCRs) is a primary determinant of β-arrestin (βarr) recruitment and trafficking. For several GPCRs such as the vasopressin receptor subtype 2 (V2R), agonist-stimulation first drives the translocation of βarrs to the plasma membrane, followed by endosomal trafficking, which is generally considered to be orchestrated by multiple phosphorylation sites. We have previously shown that mutation of a single phosphorylation site in the V2R (i.e., V2RT360A) results in near-complete loss of βarr translocation to endosomes despite robust recruitment to the plasma membrane, and compromised ERK1/2 activation. Here, we discover that a synthetic intrabody (Ib30), which selectively recognizes activated βarr1, efficiently rescues the endosomal trafficking of βarr1 and ERK1/2 activation for V2RT360A. Molecular dynamics simulations reveal that Ib30 enriches active-like βarr1 conformation with respect to the inter-domain rotation, and cellular assays demonstrate that it also enhances βarr1-β2-adaptin interaction. Our data provide an experimental framework to positively modulate the receptor-transducer-effector axis for GPCRs using intrabodies, which can be potentially integrated in the paradigm of GPCR-targeted drug discovery. © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess G protein-coupled receptors Hormone receptors Allosteric modulation of GPCR-induced β-arrestin trafficking and signaling by a synthetic intrabody info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion