TY - JOUR
T1 - Regulation of Absorption and Emission in a Protein/Fluorophore Complex
AU - Santos, Elizabeth M.
AU - Chandra, Ishita
AU - Assar, Zahra
AU - Sheng, Wei
AU - Ghanbarpour, Alireza
AU - Bingham, Courtney
AU - Vasileiou, Chrysoula
AU - Geiger, James H.
AU - Borhan, Babak
N1 - Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society
PY - 2024/8/16
Y1 - 2024/8/16
N2 - Human cellular retinol binding protein II (hCRBPII) was used as a protein engineering platform to rationally regulate absorptive and emissive properties of a covalently bound fluorogenic dye. We demonstrate the binding of a thio-dapoxyl analog via formation of a protonated imine between an active site lysine residue and the chromophore’s aldehyde. Rational manipulation of the electrostatics of the binding pocket results in a 204 nm shift in absorption and a 131 nm shift in emission. The protein is readily expressed in mammalian systems and binds with exogenously delivered fluorophore as demonstrated by live-cell imaging experiments.
AB - Human cellular retinol binding protein II (hCRBPII) was used as a protein engineering platform to rationally regulate absorptive and emissive properties of a covalently bound fluorogenic dye. We demonstrate the binding of a thio-dapoxyl analog via formation of a protonated imine between an active site lysine residue and the chromophore’s aldehyde. Rational manipulation of the electrostatics of the binding pocket results in a 204 nm shift in absorption and a 131 nm shift in emission. The protein is readily expressed in mammalian systems and binds with exogenously delivered fluorophore as demonstrated by live-cell imaging experiments.
UR - http://www.scopus.com/inward/record.url?scp=85199544401&partnerID=8YFLogxK
U2 - 10.1021/acschembio.4c00125
DO - 10.1021/acschembio.4c00125
M3 - Article
C2 - 39046136
AN - SCOPUS:85199544401
SN - 1554-8929
VL - 19
SP - 1725
EP - 1732
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 8
ER -