Reversing thromboxane A2 receptor activity from calcium to cAMP signaling by shifting Gαq to Gαs covalently linked to the receptor

Qun Ying Li, Yan Li, Asuka Inoue, Renzhong Lu, Anna Xu, Ke He Ruan

Research output: Contribution to journalArticlepeer-review


The conversion of a G-protein coupling receptor (GPCR) signaling from one G-protein to another could be a novel concept for drug designs. Thromboxane A2 (TXA2) receptor (TP) plays a key role in mediating platelet aggregation and thrombosis through its binding to TXA2 and coupling with Gαq-calcium signaling. To alter TP coupling from Gαq to Gαs, which mediates the opposite anti-platelet aggregation and vasodilation activities, we first created a recombinant single-chain (SC) TP-Gαq complex (SC-TP-Gαq) in which the Gαq N-terminus is covalently linked to the TP C-terminus. The SC-TP-Gαq expressed on HEK293 cells could perform identical functions of wild-type TP binding to agonist and activating Gαq triggering Ca2+ signal. The results were further confirmed by expressing the SC-TP-Gαq on CRISPR-edited Gαq-knocked-out HEK293 cells. A step further, by replacing Gαq using Gαs for SC-TP-Gαq, a second recombinant SC-TP-Gαs was created and expressed on wild type and CRISPR-edited Gαs-knocked-out HEK293 cells. Upon the binding of the same TXA2 agonist to the cells expressing SC-TP-Gαs, cAMP signaling was observed. The results have provided strong evidence that covalently linking TP with different Gα could control cell signaling. This study has opened a door to developing a method to redirect TP signaling from thrombotic calcium to anti-thrombotic cAMP, and apply it to that of other GPCRs.

Original languageEnglish
Article number108465
JournalBiochemical Engineering Journal
Publication statusPublished - 2022 Jun


  • Anti-platelet aggregation
  • G-protein coupling receptors (GPCR)
  • Single-chain (SC) TP-Gαq protein
  • Thromboxane A2 (TXA2) receptor
  • Vasodilation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Environmental Engineering
  • Biomedical Engineering


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