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Peptide microarrays for the detection of molecular interactions in cellular signal transduction

2005; Wiley; Volume: 5; Issue: 8 Linguagem: Inglês

10.1002/pmic.200401095

1615-9861

Oda Stoevesandt, Martin Elbs, Karsten Köhler, Annemarie C. Lellouch, Rainer Fischer, Thierry André, Roland Brock,

Glycosylation and Glycoproteins Research

The formation of protein complexes is a hallmark of cellular signal transduction. Here, we show that peptide microarrays provide a robust and quantitative means to detect signalling-dependent changes of molecular interactions. Recruitment of a protein into a complex upon stimulation of a cell leads to the masking of an otherwise exposed binding site. In cell lysates this masking can be detected by reduced binding to a microarray carrying a peptide that corresponds to the binding motif of the respective interaction domain. The method is exemplified for the lymphocyte-specific tyrosine kinase 70 kDa zeta-associated protein binding to a bis-phosphotyrosine-motif of the activated T-cell receptor via its tandem SH2 domain. Compared to established techniques, the method provides a significant shortcut to the detection of molecular interactions.