Fusion with Promiscuous G alpha(16) Subunit Reveals Signaling Bias at Muscarinic Receptors

0546460 - FGÚ 2022 RIV CH eng J - Journal Article
Randáková, Alena - Nelic, Dominik - Hochmalová, Martina - Zimčík, Pavel - Mulenga, Mutale Jane - Boulos, J. - Jakubík, Jan
Fusion with Promiscuous G alpha(16) Subunit Reveals Signaling Bias at Muscarinic Receptors.
International Journal of Molecular Sciences. Roč. 22, č. 18 (2021), č. článku 10089. E-ISSN 1422-0067
R&D Projects: GA ČR(CZ) GJ19-06106Y
Institutional support: RVO:67985823
Keywords : muscarinic receptors * signaling bias * fusion proteins * non-canonical signaling
OECD category: Pharmacology and pharmacy
Impact factor: 6.208, year: 2021
Method of publishing: Open access
https://www.mdpi.com/1422-0067/22/18/10089

A complex evaluation of agonist bias at G-protein coupled receptors at the level of G-protein classes and isoforms including non-preferential ones is essential for advanced agonist screening and drug development. Molecular crosstalk in downstream signaling and a lack of sufficiently sensitive and selective methods to study direct coupling with G-protein of interest complicates this analysis. We performed binding and functional analysis of 11 structurally different agonists on prepared fusion proteins of individual subtypes of muscarinic receptors and non-canonical promiscuous alpha-subunit of G(16) protein to study agonist bias. We have demonstrated that fusion of muscarinic receptors with G alpha(16) limits access of other competitive G alpha subunits to the receptor, and thus enables us to study activation of G alpha(16) mediated pathway more specifically. Our data demonstrated agonist-specific activation of G(16) pathway among individual subtypes of muscarinic receptors and revealed signaling bias of oxotremorine towards G alpha(16) pathway at the M-2 receptor and at the same time impaired G alpha(16) signaling of iperoxo at M-5 receptors. Our data have shown that fusion proteins of muscarinic receptors with alpha-subunit of G-proteins can serve as a suitable tool for studying agonist bias, especially at non-preferential pathways.
Permanent Link: http://hdl.handle.net/11104/0322964