Design of an engineered Gαq protein for high-yield expression and NMR isotope labeling in E. coli.
- 1State Key Laboratory of Magnetic Resonance Spectroscopy and Imaging, National Center for Magnetic Resonance in Wuhan, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, 430071, China.
- 2University of Chinese Academy of Sciences, Beijing, 100049, China.
Abstract
G protein-coupled receptors (GPCRs) are pivotal therapeutic targets that signal through heterotrimeric G proteins. While structural insights into Gαs and Gαi/o subfamilies have advanced significantly, characterizing the Gαq/11 subfamily remains challenging due to poor soluble expression in prokaryotic systems, precluding high-yield isotope labeling needed for NMR studies. Here, we present an efficient expression strategy for Gαq protein in Escherichia coli. We identified the H5 helix as a critical determinant of folding and solubility. By swapping this helix with its Gαi/o counterpart and employing low-temperature co-expression with bacterial chaperones, we significantly enhance soluble yields while retaining GTPase activity. The ¹⁵N-labeled protein exhibited well-dispersed ¹H-¹⁵N HSQC spectra, validating the suitability of this expression strategy for future high-resolution NMR investigations.
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BibTeX
@article{zhang2026design,
title = {{Design of an engineered Gαq protein for high-yield expression and NMR isotope labeling in E. coli.}},
author = {Qiang Zhang and Ju Yang and Rong Yan and Yunfei Hu},
journal = {Magnetic resonance letters},
year = {2026},
month = oct,
doi = {10.1016/j.mrl.2026.200313},
url = {https://europepmc.org/article/MED/42927389},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Design of an engineered Gαq protein for high-yield expression and NMR isotope labeling in E. coli.
AU - Qiang Zhang
AU - Ju Yang
AU - Rong Yan
AU - Yunfei Hu
PY - 2026
DA - 2026/10/16
JO - Magnetic resonance letters
DO - 10.1016/j.mrl.2026.200313
UR - https://europepmc.org/article/MED/42927389
AB - G protein-coupled receptors (GPCRs) are pivotal therapeutic targets that signal through heterotrimeric G proteins. While structural insights into Gαs and Gαi/o subfamilies have advanced significantly, characterizing the Gαq/11 subfamily remains challenging due to poor soluble expression in prokaryotic systems, precluding high-yield isotope labeling needed for NMR studies. Here, we present an efficient expression strategy for Gαq protein in Escherichia coli. We identified the H5 helix as a critical determinant of folding and solubility. By swapping this helix with its Gαi/o counterpart and employing low-temperature co-expression with bacterial chaperones, we significantly enhance soluble yields while retaining GTPase activity. The ¹⁵N-labeled protein exhibited well-dispersed ¹H-¹⁵N HSQC spectra, validating the suitability of this expression strategy for future high-resolution NMR investigations.
SN - 2097-0048
SN - 2772-5162
ER -