An Excelled Trifluorinated Probe for GPCR Conformational Quantification
- 1University of South Florida
Abstract
G protein-coupled receptors (GPCRs) represent the largest membrane protein family in the human body and the major targets of FDA-approved medications. Unlike some other physiological proteins, GPCRs are highly plastic and often exist as collections of conformational ensembles in the environment to exert their function. Therefore, understanding how a GPCR adopts different conformational ensembles in response to ligands and signaling partners is essential for drug development. 19F-NMR has emerged as one of the most promising tools in this regard; however, a well-established 19F-labeling system is still lacking, including an effective 19F probe that meets both environmental susceptibility and NMR sensitivity requirements, even though progress has been made dramatically in recent years. Thus, this manuscript reports a novel conformational probe (4-(bromomethyl)-ethoxy-2-(trifluoromethyl)benzene, BTMB) that offers paramount benefits for future GPCR conformational dynamics studies. To rationalize the mechanism behind BTMB's superiority for future probe development, we performed MD simulations, referenced to the current state-of-the-art probe, BTFMA. Our study suggests that the spatial proximity of aromatic residues significantly influences the chemical shift dispersion and linewidths of related 19F-probe-profiled conformational states, as well as the rigidity and compactness of -CF3 in the target protein.
Related
- Designing rigid protein fiducials to visualize GPCR conformational states
- Paired-probe chemoproteomic strategy enables global site-resolved quantification of lysine redox states
- Targeted degradation of a constitutively active GPCR oncoprotein
- β-arrestin1 directly engages Gαs to sustain endosomal GPCR signaling
- A co-evolved peptide-GPCR system senses host entry to drive fungal infection
BibTeX
@misc{wang2026excelled,
title = {{An Excelled Trifluorinated Probe for GPCR Conformational Quantification}},
author = {X. Wang and Y. Cao and W. Zhao and W. Sun and T. Ma and Y. Zhang and L. Ye},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.18.752616},
url = {https://www.biorxiv.org/content/10.64898/2026.09.18.752616v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - An Excelled Trifluorinated Probe for GPCR Conformational Quantification
AU - X. Wang
AU - Y. Cao
AU - W. Zhao
AU - W. Sun
AU - T. Ma
AU - Y. Zhang
AU - L. Ye
PY - 2026
DA - 2026/09/18
PB - bioRxiv
DO - 10.64898/2026.09.18.752616
UR - https://www.biorxiv.org/content/10.64898/2026.09.18.752616v1
AB - G protein-coupled receptors (GPCRs) represent the largest membrane protein family in the human body and the major targets of FDA-approved medications. Unlike some other physiological proteins, GPCRs are highly plastic and often exist as collections of conformational ensembles in the environment to exert their function. Therefore, understanding how a GPCR adopts different conformational ensembles in response to ligands and signaling partners is essential for drug development. 19F-NMR has emerged as one of the most promising tools in this regard; however, a well-established 19F-labeling system is still lacking, including an effective 19F probe that meets both environmental susceptibility and NMR sensitivity requirements, even though progress has been made dramatically in recent years. Thus, this manuscript reports a novel conformational probe (4-(bromomethyl)-ethoxy-2-(trifluoromethyl)benzene, BTMB) that offers paramount benefits for future GPCR conformational dynamics studies. To rationalize the mechanism behind BTMB's superiority for future probe development, we performed MD simulations, referenced to the current state-of-the-art probe, BTFMA. Our study suggests that the spatial proximity of aromatic residues significantly influences the chemical shift dispersion and linewidths of related 19F-probe-profiled conformational states, as well as the rigidity and compactness of -CF3 in the target protein.
ER -