Photo-SNAP-tag, a Light-Regulated Chemical Labeling System
- 16 July 2020
- journal article
- research article
- Published by American Chemical Society (ACS) in ACS Chemical Biology
- Vol. 15 (8) , 2212-2220
Abstract
Methods that allow labeling and tracking of proteins have been instrumental for understanding their function. Traditional methods for labeling proteins include fusion to fluorescent proteins or self-labeling chemical tagging systems such as SNAP-tag or Halo-tag. These latter approaches allow bright fluorophores or other chemical moieties to be attached to a protein of interest through a small fusion tag. In this work, we sought to improve the versatility of self-labeling chemical-tagging systems by regulating their activity with light. We used light-inducible dimerizers to reconstitute a split SNAP-tag (modified human O6-alkylguanine-DNA-alkyltransferase, hAGT) protein, allowing tight light-dependent control of chemical labeling. In addition, we generated a small split SNAP-tag fragment that can efficiently self-assemble with its complement fragment, allowing high labeling efficacy with a small tag. We envision these tools will extend the versatility and utility of the SNAP-tag chemical system for protein labeling applications.Keywords
Funding Information
- National Institute of General Medical Sciences (GM100225, GM136367)
This publication has 50 references indexed in Scilit:
- Photoswitchable Fluorescent Proteins: Do Not Always Look on the Bright SideACS Nano, 2016
- Quantitative assessment of fluorescent proteinsNature Methods, 2016
- Phototransformable fluorescent proteins: Future challengesCurrent Opinion in Chemical Biology, 2014
- Development of Cysteine-Free Fluorescent Proteins for the Oxidative EnvironmentPLOS ONE, 2012
- HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein AnalysisACS Chemical Biology, 2008
- An Engineered Protein Tag for Multiprotein Labeling in Living CellsChemistry & Biology, 2008
- Formation of stacked ER cisternae by low affinity protein interactionsJournal of Cell Biology, 2003
- A general method for the covalent labeling of fusion proteins with small molecules in vivoNature Biotechnology, 2002
- A Photoactivatable GFP for Selective Photolabeling of Proteins and CellsScience, 2002
- Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live CellsScience, 2002