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

ABSTRACT: 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 (ΔF/F _min ≈ 18) designated Citroff1 and a direct-response biosensor

SUBMITTER: Zhao Y 

PROVIDER: S-EPMC7453566 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature