Back to Search
Start Over
Simultaneous triggering of protein activity and fluorescence.
- Source :
-
Journal of the American Chemical Society [J Am Chem Soc] 2004 Jun 16; Vol. 126 (23), pp. 7170-1. - Publication Year :
- 2004
-
Abstract
- Many areas of biology can benefit greatly from methods to spatially and temporally control protein activity. Here, we describe an approach that allows the simultaneous photo-triggering of the activity and the fluorescence of a protein. Smad2, a protein central to the transforming growth factor-beta (TGF-beta) signal transduction pathway, was modified with a fluorophore and a photocleavable moiety that acted as both a caging and a fluorescence quenching group. In its caged state, the protein formed a non-fluorescent heterodimer with the protein SARA. Irradiation with UV light and photocleavage of the caging group produced a fluorescent homotrimer. These in vitro experiments demonstrated that a photochemical trigger mimicking the critical biochemical event of serine phosphorylation involved in the TGF-beta signaling pathway could be obtained and that fluorescence could be used as a read-out of protein activity. This approach should prove particularly useful for the monitoring of a protein's activity and location inside of living cells.
- Subjects :
- Chromatography, Gel
Chromatography, High Pressure Liquid
DNA-Binding Proteins chemical synthesis
DNA-Binding Proteins chemistry
Fluorescence
Molecular Structure
Smad2 Protein
Spectrometry, Fluorescence
Trans-Activators chemical synthesis
Trans-Activators chemistry
DNA-Binding Proteins analysis
DNA-Binding Proteins metabolism
Trans-Activators analysis
Trans-Activators metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0002-7863
- Volume :
- 126
- Issue :
- 23
- Database :
- MEDLINE
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 15186142
- Full Text :
- https://doi.org/10.1021/ja0499142