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Gγ and Gα Identity Dictate a G-Protein Heterotrimer Plasma Membrane Targeting

Mystek, Paweł and Rysiewicz, Beata and Gregrowicz, Jan and Dziedzicka-Wasylewska, Marta and Polit, Agnieszka (2019) Gγ and Gα Identity Dictate a G-Protein Heterotrimer Plasma Membrane Targeting. Cells, 8 (10). Art. No. 1246. ISSN 2073-4409. https://resolver.caltech.edu/CaltechAUTHORS:20191022-100603148

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Abstract

Heterotrimeric G-proteins along with G-protein-coupled receptors (GPCRs) regulate many biochemical functions by relaying the information from the plasma membrane to the inside of the cell. The lipid modifications of Gα and Gγ subunits, together with the charged regions on the membrane interaction surface, provide a peculiar pattern for various heterotrimeric complexes. In a previous study, we found that Gαs and Gαi3 prefer different types of membrane-anchor and subclass-specific lipid domains. In the present report, we examine the role of distinct Gγ subunits in the membrane localization and spatiotemporal dynamics of Gαs and Gαi3 heterotrimers. We characterized lateral diffusion and G-protein subunit interactions in living cells using fluorescence recovery after photobleaching (FRAP) microscopy and fluorescence resonance energy transfer (FRET) detected by fluorescence lifetime imaging microscopy (FLIM), respectively. The interaction of Gγ subunits with specific lipids was confirmed, and thus the modulation of heterotrimeric G-protein localization. However, the Gα subunit also modulates trimer localization, and so the membrane distribution of heterotrimeric G-proteins is not dependent on Gγ only.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.3390/cells8101246DOIArticle
ORCID:
AuthorORCID
Polit, Agnieszka0000-0001-5811-9838
Additional Information:© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). Received: 17 September 2019; Accepted: 11 October 2019; Published: 13 October 2019. Author Contributions: Conceptualization: A.P. and P.M. Formal analysis: A.P., P.M. and B.R. Funding acquisition: A.P. Investigation: Genetics constructs: P.M. and J.G. FRAP experiments: P.M. FLIM–FRET experiments: A.P. and J.G. cAMP production experiments: B.R. Pull-down assay: A.P. and P.M. Project administration: A.P. Visualization: A.P., P.M. and B.R. Writing—original draft: A.P. and P.M. Writing—review and editing: A.P., M.D.-W. and P.M. This work was supported by a grant awarded by Polish National Center for Science (NCN) no. 2016/23/B/NZ1/00530. The authors would like to thank Jerzy Dobrucki—the head of the Department of Cell Biophysics of Jagiellonian University for providing access to the confocal microscopes. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
National Science Centre (Poland)2016/23/B/NZ1/00530
Subject Keywords:protein-membrane interaction; spatiotemporal protein dynamics; G-proteins; FLIM-FRET; FRAP
Issue or Number:10
Record Number:CaltechAUTHORS:20191022-100603148
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191022-100603148
Official Citation:Mystek, P.; Rysiewicz, B.; Gregrowicz, J.; Dziedzicka-Wasylewska, M.; Polit, A. Gγ and Gα Identity Dictate a G-Protein Heterotrimer Plasma Membrane Targeting. Cells 2019, 8, 1246
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99385
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:22 Oct 2019 20:21
Last Modified:22 Oct 2019 20:21

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