Fluorogenic Protein Probes with Red and Near-Infrared Emission for Genetically Targeted Imaging

被引：16

作者：

Bachollet, Sylvestre P. J. T. ^{[1
]}

Addi, Cyril ^{[2
]}

Pietrancosta, Nicolas ^{[1
,3
]}

Mallet, Jean-Maurice ^{[1
]}

Dumat, Blaise ^{[1
]}

机构：

[1] Sorbonne Univ, PSL Univ, CNRS, Lab Biomol,Dept Chim,Ecole Normale Super,LBM, F-75005 Paris, France

[2] Sorbonne Univ, Inst Pasteur, CNRS UMR3691, Cell Biol & Infect Dept,Membrane Traff & Cell Div, F-75005 Paris, France

[3] Sorbonne Univ, INSERM, CNRS, Neurosci Paris Seine,Inst Biol Paris Seine NPS IB, Paris, France

来源：

CHEMISTRY-A EUROPEAN JOURNAL | 2020年 / 26卷 / 63期

关键词：

fluorescent probes; HaloTag; imaging agents; molecular dynamics; molecular rotors; LABELING IN-VITRO; LIVE-CELL; TETRAZINE PROBES; FUSION PROTEINS; GENERAL-METHOD; SERUM-ALBUMIN; DYES; TAG; DESIGN; TECHNOLOGY;

D O I：

10.1002/chem.202002911

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Fluorogenic probes are important tools to image proteins with high contrast and no wash protocols. In this work, we rationally designed and synthesized a small set of four protein fluorogens with red or near-infrared emission. The fluorophores were characterized in the presence of albumin as a model protein environment and exhibited good fluorogenicity and brightness (fluorescence quantum yield up to 36%). Once conjugated to a haloalkane ligand, the probes reacted with the protein self-labeling tag HaloTag with a high fluorescence enhancement (up to 156-fold). The spectroscopic properties of the fluorogens and their reaction with HaloTag were investigated experimentally in vitro and with the help of molecular dynamics. The two most promising probes, one in the red and one in the near-infrared range, were finally applied to image the nucleus or actin in live-cell and in wash-free conditions using fluorogenic and chemogenetic targeting of HaloTag fusion proteins.

引用

页码：14467 / 14473

页数：7

共 51 条

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