High-Performance Intensiometric Direct- and Inverse-Response Genetically Encoded Biosensors for Citrate

• DOI: 10.1021/acscentsci.0c00518
• Publication Date: 2020-07-09T20:43:33Z
• Authors: Yufeng Zhao; Yi Shen; Yurong Wen; Robert E. Campbell
• Keywords: Chemistry; 0303 health sciences; 03 medical and health sciences; QD1-999

Motivated by the growing recognition of citrate as a central metabolite in variety biological processes associated with healthy and diseased cellular states, we have developed series high-performance genetically encoded biosensors suitable for imaging concentrations mammalian cells. The design these was guided structural studies citrate-responsive sensor histidine kinase took advantage same conformational changes proposed to propagate from binding domain catalytic domain. Following extensive engineering based on combination structure mutagenesis directed evolution, produced an inverse-response biosensor (ΔF/Fmin ≈ 18) designated Citroff1 direct-response 9) Citron1. We report X-ray crystal Citron1 demonstrate utility both qualitative quantitative steady-state pharmacologically perturbed live