Regulation of Absorption and Emission in a Protein/Fluorophore Complex

Overview

Journal ACS Chem Biol

Publisher American Chemical Society

Specialties Biochemistry
Biology

Date 2024 Jul 24

PMID 39046136

Authors

Elizabeth M Santos

Ishita Chandra

Zahra Assar

Wei Sheng

Alireza Ghanbarpour

Courtney Bingham

Chrysoula Vasileiou

James H Geiger

Babak Borhan

Affiliations

Soon will be listed here.

Abstract

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.

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