Fluorogenic Protein Probes with Red and Near-Infrared Emission for Genetically Targeted Imaging*.
Chemistry
; 26(63): 14467-14473, 2020 Nov 11.
Article
em En
| MEDLINE
| ID: mdl-32691883
ABSTRACT
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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Corantes Fluorescentes
/
Raios Infravermelhos
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article