Investigation of the effect of GPCR oligomerization on the GNAi1 protein homodimerization in live cells using FRET

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2022-1-26

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Nalli, Enise

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G-Protein Coupled Receptors (GPCR) are membrane proteins that pass the cell membrane seven times. In classical GPCR signaling pathways, one GPCR-one heterotrimeric G-protein interaction model is enough to transmit the signal to effector proteins. Studies since 2000 showed that one GPCR dimer-one heterotrimeric G-protein interaction model is more likely, and GPCRs having homo- /hetero- dimers interact with a single G⍺-protein. Recently, studies on GPCRs indicated that more than two receptors interact to form active receptor oligomers during signal transduction. Navarro et al. showed that within a heterotetrameric receptor complex, formed by the dimerization of the dimers, the G proteins interacting with the dimers were brought into close proximity (Navarro et al., 2018). Furthermore, studies with Ras proteins, which are members of the G-protein family, have shown that these proteins form dimers playing important roles in various signaling pathways. More recently, a member of our Lab., Özge Atay, has shown the physical interaction of Gαi1 proteins on the cell membrane. However, it is still not vi clear whether the Gαi1 protein homodimerization is a result of the formation of receptor tetramers or if the Gαi1 homodimers form independently of the receptors. Receptor independent G-protein dimerization might play a role in stabilizing the receptor tetramers. In order to answer this question, Förster Resonance Energy Transfer (FRET) method was used to quantitatively investigate the effect of GPCR oligomerization on the Gαi1 homodimerization under two conditions: (1) blocking GPCR-Gαi1 interaction with Gαi1-specific minigenes and (2) receptor oligomerization by agonist (Quinpirole) treatment.

Subject Keywords

Gαi1 protein, Homodimer formation, Receptor oligomerization, Receptor dependency, FRET

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https://hdl.handle.net/11511/96067

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Graduate School of Natural and Applied Sciences, Thesis

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Citation Formats

E. Nalli, “Investigation of the effect of GPCR oligomerization on the GNAi1 protein homodimerization in live cells using FRET,” M.S. - Master of Science, Middle East Technical University, 2022.