G-Protein-Coupled Receptors: Membrane Diffusion and Organization Matter

Haanappel, Evert; Salomé, Laurence

doi:10.1007/978-3-319-66601-3_11

Cited by 2 publications

(2 citation statements)

References 107 publications

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“…As a result, single-molecule based approaches enable observation of heterogeneous behavior (and transient phe-nomena) within subpopulations of molecules (43). Singlemolecule techniques such as single particle tracking (SPT) have emerged as a powerful approach to explore the organization and lateral diffusion of membrane-bound molecules (31,42,44,45). Although SPT has been used to probe dynamics of membrane-bound molecules (39,46), the application of SPT to GPCRs is relatively rare (47,48).…”

Section: Introductionmentioning

confidence: 99%

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Shrivastava

Sarkar

Preira

et al. 2020

Biophysical Journal

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G protein-coupled receptors (GPCRs) are important membrane proteins in higher eukaryotes that carry out a vast array of cellular signaling and act as major drug targets. The serotonin 1A receptor is a prototypical member of the GPCR family and is implicated in neuropsychiatric disorders such as anxiety and depression, besides serving as an important drug target. With an overall goal of exploring the functional consequence of altered receptor dynamics, in this work, we probed the role of the actin cytoskeleton in the dynamics, ligand binding, and signaling of the serotonin 1A receptor. We monitored receptor dynamics utilizing single particle tracking, which provides information on relative distribution of receptors in various diffusion modes in addition to diffusion coefficient. We show here that the short-term diffusion coefficient of the receptor increases upon actin destabilization by cytochalasin D. In addition, analysis of individual trajectories shows that there are changes in relative populations of receptors undergoing various types of diffusion upon actin destabilization. The release of dynamic constraint was evident by an increase in the radius of confinement of the receptor upon actin destabilization. The functional implication of such actin destabilization was manifested as an increase in specific agonist binding and downstream signaling, monitored by measuring reduction in cellular cAMP levels. These results bring out the interdependence of GPCR dynamics with cellular signaling. CD treatment of cellsActin cytoskeleton destabilization in HEK-293 cells expressing the myctagged serotonin 1A receptor was performed utilizing CD as described earlier (26,50,51) with few modifications. Briefly, 2 mM CD stock solution was prepared in DMSO and working stock was made upon diluting this stock in serum-free DMEM/F-12 medium. Cells were grown in complete Serotonin 1A Receptor Dynamics & Function

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Section: Introductionmentioning

confidence: 99%

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Shrivastava

Sarkar

Preira

et al. 2020

Biophysical Journal

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“…SPT has emerged as a powerful approach to monitor lateral diffusion and organization of membrane lipids and proteins. ,,,,, Although single particle tracking has been utilized to measure dynamics of membrane proteins, , the application of SPT to GPCRs is relatively limited. ,,, In this work, we explored the diffusion characteristics of the human serotonin 1A receptor in live cells under acute and chronic cholesterol depleted conditions utilizing SPT. Analysis of GPCR diffusion in the membrane helps understand GPCR function and provides insight into cellular signaling .…”

Section: Introductionmentioning

confidence: 99%

Cholesterol-Dependent Dynamics of the Serotonin_1A Receptor Utilizing Single Particle Tracking: Analysis of Diffusion Modes

et al. 2022

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G protein-coupled receptors (GPCRs) are signaling hubs in cell membranes that regulate a wide range of physiological processes and are popular drug targets. Serotonin1A receptors are important members of the GPCR family and are implicated in neuropsychiatric disorders. Cholesterol is a key constituent of higher eukaryotic membranes and is believed to contribute to the segregated distribution of membrane constituents into domains. To explore the role of cholesterol in lateral dynamics of GPCRs, we utilized single particle tracking (SPT) to monitor diffusion of serotonin1A receptors under acute and chronic cholesterol-depleted conditions. Our results show that the short-term diffusion coefficient of the receptor decreases upon cholesterol depletion, irrespective of the method of cholesterol depletion. Analysis of SPT trajectories revealed that relative populations of receptors undergoing various modes of diffusion change upon cholesterol depletion. Notably, in cholesterol-depleted cells, we observed an increase in the confined population of the receptor accompanied by a reduction in diffusion coefficient for chronic cholesterol depletion. These results are supported by our recent work and present observations that show polymerization of G-actin in response to chronic cholesterol depletion. Taken together, our results bring out the interdependence of cholesterol and actin cytoskeleton in regulating diffusion of GPCRs in membranes.

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G-Protein-Coupled Receptors: Membrane Diffusion and Organization Matter

Cited by 2 publications

References 107 publications

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Cholesterol-Dependent Dynamics of the Serotonin_1A Receptor Utilizing Single Particle Tracking: Analysis of Diffusion Modes

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G-Protein-Coupled Receptors: Membrane Diffusion and Organization Matter

Cited by 2 publications

References 107 publications

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Role of Actin Cytoskeleton in Dynamics and Function of the Serotonin1A Receptor

Cholesterol-Dependent Dynamics of the Serotonin1A Receptor Utilizing Single Particle Tracking: Analysis of Diffusion Modes

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Cholesterol-Dependent Dynamics of the Serotonin_1A Receptor Utilizing Single Particle Tracking: Analysis of Diffusion Modes