Monitoring TRPC7 Conformational Changes by BRET Following GPCR Activation

Pétigny, Cécile; Dumont, Audrey-Ann; Giguère, Hugo; Collette, Audrey; Holleran, Brian J.; Iftinca, Mircea; Altier, Christophe; Besserer-Offroy, Élie; Auger‐Messier, Mannix; Leduc, Richard

doi:10.3390/ijms23052502

Cited by 2 publications

(2 citation statements)

References 52 publications

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“…Figure 1A shows that there is a 2.5% signal increase in the group stimulated by magnetic field over the non-stimulated group. The response seen is comparable to other BRET studies of single-protein conformational changes ( Audet et al, 2008 ; Pétigny et al, 2022 ). We then designed a BRET construct to test if the protein underwent a dimerization event due to magnetic stimulus.…”

Section: Resultssupporting

confidence: 87%

A putative design for the electromagnetic activation of split proteins for molecular and cellular manipulation

Grady,

Castellanos Franco,

Schossau

et al. 2024

Front. Bioeng. Biotechnol.

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The ability to manipulate cellular function using an external stimulus is a powerful strategy for studying complex biological phenomena. One approach to modulate the function of the cellular environment is split proteins. In this method, a biologically active protein or an enzyme is fragmented so that it reassembles only upon a specific stimulus. Although many tools are available to induce these systems, nature has provided other mechanisms to expand the split protein toolbox. Here, we show a novel method for reconstituting split proteins using magnetic stimulation. We found that the electromagnetic perceptive gene (EPG) changes conformation due to magnetic field stimulation. By fusing split fragments of a certain protein to both termini of the EPG, the fragments can be reassembled into a functional protein under magnetic stimulation due to conformational change. We show this effect with three separate split proteins: NanoLuc, APEX2, and herpes simplex virus type-1 thymidine kinase. Our results show, for the first time, that reconstitution of split proteins can be achieved only with magnetic fields. We anticipate that this study will be a starting point for future magnetically inducible split protein designs for cellular perturbation and manipulation. With this technology, we can help expand the toolbox of the split protein platform and allow better elucidation of complex biological systems.

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Section: Resultssupporting

confidence: 87%

A putative design for the electromagnetic activation of split proteins for molecular and cellular manipulation

Grady,

Castellanos Franco,

Schossau

et al. 2024

Front. Bioeng. Biotechnol.

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“…As for TRPC7 specifically, the conformational changes observed in the TRPC7 channel in response to angiotensin II stimulation in rat fibroblasts imply a role for the channel in fibrosis ( Petigny et al, 2022 ). However, the TRPC7 protein is not detectable in human fibroblasts ( Ikeda et al, 2013 ) and its mRNA is not altered in response to profibrotic endothelin-1 stimulation in rat neonatal fibroblasts ( Nishida et al, 2007 ), hence making questionable the role of TRPC7.…”

Section: Trp Channel Expression and Roles In Cardiac Fibroblastsmentioning

confidence: 99%

Emerging role of transient receptor potential (TRP) ion channels in cardiac fibroblast pathophysiology

Gwanyanya

Mubagwa

2022

Front. Physiol.

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Cardiac fibroblasts make up a major proportion of non-excitable cells in the heart and contribute to the cardiac structural integrity and maintenance of the extracellular matrix. During myocardial injury, fibroblasts can be activated to trans-differentiate into myofibroblasts, which secrete extracellular matrix components as part of healing, but may also induce cardiac fibrosis and pathological cardiac structural and electrical remodeling. The mechanisms regulating such cellular processes still require clarification, but the identification of transient receptor potential (TRP) channels in cardiac fibroblasts could provide further insights into the fibroblast-related pathophysiology. TRP proteins belong to a diverse superfamily, with subgroups such as the canonical (TRPC), vanilloid (TRPV), melastatin (TRPM), ankyrin (TRPA), polycystin (TRPP), and mucolipin (TRPML). Several TRP proteins form non-selective channels that are permeable to cations like Na+ and Ca2+ and are activated by various chemical and physical stimuli. This review highlights the role of TRP channels in cardiac fibroblasts and the possible underlying signaling mechanisms. Changes in the expression or activity of TRPs such as TRPCs, TRPVs, TRPMs, and TRPA channels modulate cardiac fibroblasts and myofibroblasts, especially under pathological conditions. Such TRPs contribute to cardiac fibroblast proliferation and differentiation as well as to disease conditions such as cardiac fibrosis, atrial fibrillation, and fibroblast metal toxicity. Thus, TRP channels in fibroblasts represent potential drug targets in cardiac disease.

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Monitoring TRPC7 Conformational Changes by BRET Following GPCR Activation

Cited by 2 publications

References 52 publications

A putative design for the electromagnetic activation of split proteins for molecular and cellular manipulation

A putative design for the electromagnetic activation of split proteins for molecular and cellular manipulation

Emerging role of transient receptor potential (TRP) ion channels in cardiac fibroblast pathophysiology

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