1. Academic Validation
  2. Coumarin-Conjugated Macromolecular Probe for Sequential Stimuli-Mediated Activation

Coumarin-Conjugated Macromolecular Probe for Sequential Stimuli-Mediated Activation

  • Bioconjug Chem. 2023 Dec 13. doi: 10.1021/acs.bioconjchem.3c00418.
Yu Jiang 1 Runqi Li 1 Fei Ren 1 Shuke Yang 1 Andong Shao 1
Affiliations

Affiliation

  • 1 School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, China.
Abstract

Protein bioconjugation has emerged as one of the most valuable tools for the development of protein-based biochemical assays. Here, we report a fluorescent macromolecular material, RF16_Halo, in which the coumarin derivative RF16 is specifically conjugated onto HaloTag protein to achieve a dual-stimuli-mediated fluorescence response. RF16 is first obtained by installing a H2O2-sensitive boron cage onto the C7 hydroxy moiety of the coumarin fluorophore with a HaloTag ligand attaching to the pH-labile 1,3-dioxane moiety. Upon stimulation, RF16_Halo exhibits a sequential fluorescence response to H2O2/pH at both liquid and solid interfaces. The fluorescence of the RF16_Halo-based protein film increases linearly toward H2O2 with a higher sensitivity when compared with that of RF16. Subsequently, the H2O2-cleaved RF16_Halo presents a pH-dependent fluorescence decrease under acidic conditions. Such a stimulus-responsive fluorescence "off-on-off" multimode enables RF16_Halo to be applied as a sequential logic circuit. In addition, we evaluate the fluorescence labeling ability of RF16 to intracellular IRE1_Halo protein and demonstrate that RF16 containing the HaloTag ligand could be precisely retained in cells to track IRE1_Halo protein. Hence, we provide a unique structural design strategy to construct a fluorescence dual-responsive macromolecular probe for information encryption and protein tracking in cells.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-162075
    荧光染料