RT Journal Article
SR Electronic
T1 Multifunctional fluorophores for live-cell imaging and affinity capture of proteins
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.07.02.498544
DO 10.1101/2022.07.02.498544
A1 Kumar, Pratik
A1 Vevea, Jason D.
A1 Chapman, Edwin R.
A1 Lavis, Luke D.
YR 2022
UL http://biorxiv.org/content/early/2022/07/03/2022.07.02.498544.abstract
AB The development of enzyme-based self-labeling tags allow the labeling of proteins in living cells with synthetic small-molecules. Use of a fluorophore-containing ligand enables the visualization of protein location inside cells using fluorescence microscopy. Alternatively, deployment of a biotin-containing ligand allows purification of tagged protein using affinity resins. Despite these various applications of self-labeling tags, most ligands serve a single purpose. Here, we describe self-labeling tag ligands that allow both visualization and subsequent capture of a protein. A key design principle is exploiting the chemical properties and size of a rhodamine fluorophore to optimize cell-permeability of the ligand and the capture efficiency of the biotin conjugate. This work generates useful “multifunctional” fluorophores with generalizable design principles that will allow the construction of new tools for biology.Competing Interest StatementThe authors have declared no competing interest.