Photoactive Yellow Protein-Based Protein Labeling System with Turn-On Fluorescence Intensity
- 30 October 2009
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 131 (46) , 16610-16611
Abstract
Protein labeling provides significant information about protein function. In this research, we developed a novel protein labeling technique by utilizing photoactive yellow protein (PYP). PYP is a small protein (14 kDa) derived from purple bacteria and binds to 7-hydroxycoumarin-3-carboxylic acid as well as to a natural ligand, 4-hydroxycinnamic acid, through a thioester bond with Cys69. Based on the structure and fluorescence property of this coumarin derivative, we designed two fluorescent probes that bind to PYP. One has an azido moiety, which allows stepwise labeling by click chemistry, and the other is a fluorogenic probe. The live-cell imaging and specific labeling of PYP were achieved by using both probes. The flexibility of the probe design and the small size of the tag protein are great advantages of this system against the existing methods. This novel labeling technique can be used in a wide variety of applications for biological research.Keywords
This publication has 20 references indexed in Scilit:
- Covalent Protein Labeling Based on Noncatalytic β-Lactamase and a Designed FRET SubstrateJournal of the American Chemical Society, 2009
- Site-specific chemical modification of recombinant proteins produced in mammalian cells by using the genetically encoded aldehyde tagProceedings of the National Academy of Sciences, 2009
- GFP Family: Structural Insights into Spectral TuningChemistry & Biology, 2008
- An Eight Residue Fragment of an Acyl Carrier Protein Suffices for Post-Translational Introduction of Fluorescent Pantetheinyl Arms in Protein Modification in vitro and in vivoJournal of the American Chemical Society, 2008
- An Engineered Protein Tag for Multiprotein Labeling in Living CellsChemistry & Biology, 2008
- Fluorescent biosensors of protein functionCurrent Opinion in Chemical Biology, 2008
- Chemical probes shed light on protein functionCurrent Opinion in Structural Biology, 2007
- A guide to choosing fluorescent proteinsNature Methods, 2005
- Site-specific labeling of proteins with small molecules in live cellsCurrent Opinion in Biotechnology, 2005
- Use of Chimeric Fluorescent Proteins and Fluorescence Resonance Energy Transfer to Monitor Cellular ResponsesCirculation Research, 2004