Abstract
Multiprotein complexes play an essential role in the propagation and integration of cellular signals. Here, peptide microarrays are employed for network-based analyses of signalling-dependent changes in molecular interactions. The peptides correspond to known interaction motifs for SH2 and SH3 domains in cellular signalling proteins. Lysates of resting or stimulated cells are incubated on these arrays and the binding of signalling proteins is detected by immunofluorescence. If, due to signalling-dependent complex formation, a binding site of a protein is blocked, the signal for this protein on the array will be reduced. In addition, complex formation may co-recruit a protein to a peptide on the array leading to the appearance of a signal. Peptides and detected proteins are selected based on available knowledge of the network. This knowledge enables a significant reduction in cell number in comparison to standard mass spectrometry-based proteomics methods for the analysis of molecular complexes. Applications comprise of a functional comparison of signalling networks in different cell types, the determination of the architecture of signalling complexes, the identification of new interactions and analyses of the role of interaction domains in connecting the network.
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Sinzinger, M.D., Brock, R. (2010). Peptide Microarrays for a Network Analysis of Changes in Molecular Interactions in Cellular Signalling. In: Choi, S. (eds) Systems Biology for Signaling Networks. Systems Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-5797-9_28
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DOI: https://doi.org/10.1007/978-1-4419-5797-9_28
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