Design of an engineered Gα_q protein for high-yield expression and NMR isotope labeling in E. coli.

Authors

Qiang Zhang, Ju Yang, Rong Yan, Yunfei Hu

Published in

Magnetic resonance letters. Volume 7. Issue 3. Pages 200313. Epub Oct 16, 2026.

Abstract

G protein-coupled receptors (GPCRs) are pivotal therapeutic targets that signal through heterotrimeric G proteins. While structural insights into Gα_s and Gα_i/o subfamilies have advanced significantly, characterizing the Gα_q/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 Gα_q protein in Escherichia coli. We identified the H5 helix as a critical determinant of folding and solubility. By swapping this helix with its Gα_i/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.

PMID:
42927389
Bibliographic data and abstract were imported from PubMed on 25 Oct 2026.

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