Gγ and Gα Identity Dictate a G-Protein Heterotrimer Plasma Membrane Targeting
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.
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.Attached Files
Published - cells-08-01246.pdf
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Additional details
- Eprint ID
- 99385
- Resolver ID
- CaltechAUTHORS:20191022-100603148
- 2016/23/B/NZ1/00530
- National Science Centre (Poland)
- Created
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2019-10-22Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field