Phospho-specific recognition by 14-3-3 proteins and antibodies monitored by a high throughput label-free optical biosensor

Meng Wu, Brian Coblitz, Sojin Shikano, Shunyou Long, Matt Spieker, Anthony G. Frutos, Sunil Mukhopadhyay, Min Li

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5681-5689
Number of pages9
JournalFEBS Letters
Volume580
Issue number24
DOIs
StatePublished - Oct 16 2006

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Keywords

  • 14-3-3 and phosphopeptide
  • High throughput
  • Label-free
  • Optical biosensor
  • Resonant waveguide grating

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Wu, M., Coblitz, B., Shikano, S., Long, S., Spieker, M., Frutos, A. G., Mukhopadhyay, S., & Li, M. (2006). Phospho-specific recognition by 14-3-3 proteins and antibodies monitored by a high throughput label-free optical biosensor. FEBS Letters, 580(24), 5681-5689. https://doi.org/10.1016/j.febslet.2006.09.019