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A Genetically Encoded Ratiometric pH Probe: Wavelength Regulation‐Inspired Design of pH Indicators
Author(s) -
Berbasova Tetyana,
Tahmasebi Nick Setare,
Nosrati Meisam,
Nossoni Zahra,
Santos Elizabeth M.,
Vasileiou Chrysoula,
Geiger James H.,
Borhan Babak
Publication year - 2018
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800050
Subject(s) - fluorophore , chemistry , fluorescence , absorption (acoustics) , biophysics , photochemistry , materials science , biology , optics , physics , composite material
Mutants of human cellular retinol‐binding protein II (hCRBPII) were engineered to bind a julolidine retinal analogue for the purpose of developing a ratiometric pH sensor. The design relied on the electrostatic influence of a titratable amino acid side chain, which affects the absorption and, thus, the emission of the protein / fluorophore complex. The ratio of emissions obtained at two excitation wavelengths that correspond to the absorption of the two forms of the protein/fluorophore complex, leads to a concentration‐independent measure of pH.