Peptide microarrays for the detection of molecular interactions in cellular signal transduction |
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Authors: | Stoevesandt Oda Elbs Martin Köhler Karsten Lellouch Annemarie C Fischer Rainer André Thomas Brock Roland |
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Institution: | Institute for Cell Biology, University of Tübingen, Germany. |
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Abstract: | 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. |
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Keywords: | Cellular immunity Fluorescence correlation spectroscopy Signal transduction domain |
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