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TitleHigh-Performance Intensiometric Direct- and Inverse-Response Genetically Encoded Biosensors for Citrate.
Publication TypeJournal Article
Year of Publication2020
AuthorsZhao, Yufeng, Yi Shen, Yurong Wen, and Robert E. Campbell
JournalACS Cent Sci
Date Published2020 Aug 26

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

Alternate JournalACS Cent Sci
PubMed ID32875085
PubMed Central IDPMC7453566