Phospho-specific recognition by 14-3-3 proteins and antibodies monitored by a high throughput label-free optical biosensor
| Autor: | Matt Spieker, Brian Coblitz, Sunil Mukhopadhyay, Sojin Shikano, Meng Wu, Anthony G. Frutos, Shunyou Long, Min Li |
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| Rok vydání: | 2006 |
| Předmět: |
Phosphopeptides
Optical biosensor Biophysics Peptide Biosensing Techniques Sensitivity and Specificity Biochemistry 03 medical and health sciences 0302 clinical medicine Structural Biology High throughput Genetics Phosphorylation Surface plasmon resonance Molecular Biology Throughput (business) 030304 developmental biology chemistry.chemical_classification 0303 health sciences Molecular Structure biology Cell Biology Small molecule Resonant waveguide grating 14-3-3 Proteins chemistry 14-3-3 and phosphopeptide biology.protein Label-free Antibody Antibodies Phospho-Specific Biosensor 030217 neurology & neurosurgery Fluorescence anisotropy Protein Binding |
| Zdroj: | FEBS Letters. 580:5681-5689 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/j.febslet.2006.09.019 |
| Popis: | Label-free detection of molecular interactions has considerable potential in facilitating assay development. When combined with high throughput capability, it may be applied to small molecule screens for drug candidates. Phosphorylation is a key posttranslational process that confers diverse regulation in biological systems involving specific protein–protein interactions recognizing the phosphorylated motifs. Using a resonant waveguide grating biosensor, the Epic™ System, we have developed a generic assay to quantitatively measure phospho-specific interactions between a trafficking signal-phosphorylated SWTY peptide and 14-3-3 proteins or anti-phosphopeptide antibodies. Compared with a solution-based fluorescence anisotropy assay, our results support that the high throughput resonant waveguide grating biosensor system has favorable technical profiles in detecting protein–protein interactions that recognize phosphorylated motifs. Hence it provides a new generic HTS platform for phospho-detection. |
| Databáze: | OpenAIRE |
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