A Genetically Encoded, Ratiometric Fluorescent Biosensor for Hydrogen Sulfide.

ABSTRACT: As an important gasotransmitter, hydrogen sulfide (H₂S) plays crucial roles in cell signaling. Incorporation of p-azidophenylalanine ( pAzF) into fluorescent proteins (FPs) via genetic code expansion has been a successful strategy in developing intensity-based, genetically encoded fluorescent biosensors for H₂S. To extend this strategy for ratiometric measurement which eliminates many detection uncertainties via self-calibration at two wavelengths, we modified the chromophore of a circularly permutated, superfolder green fluorescent protein (cpsGFP) with pAzF to derive cpsGFP- pAzF, which subsequently served as a Förster resonance energy transfer (FRET) acceptor to EBFP2, an enhanced blue fluorescent protein. The resultant construct, namely, hsFRET, is the first ratio

SUBMITTER: Youssef S 

PROVIDER: S-EPMC6599572 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature