Multiplexed Detection of Protein−Peptide Interaction and Inhibition Using Capillary Electrophoresis

• Published in: Analytical chemistry (Washington) Vol. 79; no. 4; pp. 1690 - 1695
• Main Authors: Yang, Peilin, Whelan, Rebecca J, Mao, Yingwei, Lee, Angel W.-M, Carter-Su, Christin, Kennedy, Robert T
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15.02.2007
• Subjects: Analytical chemistry; Anisotropy; Binding sites; Biochemistry; Chemistry; Chromatographic methods and physical methods associated with chromatography; Electrophoresis, Capillary - instrumentation; Electrophoresis, Capillary - methods; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Exact sciences and technology; Fluorescence; Molecular Structure; Molecular weight; Other chromatographic methods; Peptides; Peptides - analysis; Peptides - antagonists & inhibitors; Protein Binding; Proteins; Proteins - analysis; Proteins - antagonists & inhibitors; Sensitivity and Specificity; Spectrometric and optical methods; Structure-Activity Relationship; Time Factors; Drug; Chemical analysis; Peptides; Capillary electrophoresis; High speed; Inhibitor; Selectivity; Inhibition; Molecular mass; Protein; Fluorescence spectrometry

High-speed capillary electrophoresis (CE) was employed to detect binding and inhibition of SH2 domain proteins using fluorescently labeled phosphopeptides as affinity probes. Single SH2 protein−phosphopeptide complexes were detected and confirmed by competition and fluorescence anisotropy. The assay...