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Phospho‐specific recognition by 14‐3‐3 proteins and antibodies monitored by a high throughput label‐free optical biosensor
Author(s) -
Wu Meng,
Coblitz Brian,
Shikano Sojin,
Long Shunyou,
Spieker Matt,
Frutos Anthony G.,
Mukhopadhyay Sunil,
Li Min
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.09.019
Subject(s) - biosensor , phosphopeptide , phosphorylation , peptide , chemistry , fluorescence anisotropy , high throughput screening , computational biology , biochemistry , biology , membrane
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.