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Fig. 1 | Cell Communication and Signaling

Fig. 1

From: High-throughput methods for identification of protein-protein interactions involving short linear motifs

Fig. 1

Schematic representation of discussed techniques for the identification of motif-based interactions. Orange represents target protein; blue hexagon represents a binding motif; yellow, green and purple represent non-binding sequences peptides. Pink star represent a detection signal e.g. fluorescence. a Peptide microarray: Peptides with known sequences are synthesized on a solid support, incubated with the target protein and interactions are detected with specific antibodies or labeled target protein. b Protein array: A selection of different purified proteins are spotted on a solid support and incubated with a labeled peptide. c Peptide phage-display: Bait protein is immobilized and used in selections against a peptide phage library. Unbound phage particles are washed away, bound phage eluted and amplified, and used for repeated rounds of selections. Enriched binding clones are sequenced. d Yeast surface display: A library of peptides are displayed on the surface of yeast cells and incubated with a target protein. The target protein is labeled with a fluorescent tag and the cells are sorted based on peptide binding using FACS. Sorted pools are sequenced. e Yeast-two-hybrid: The binding domain (BD) of a transcription factor is linked to the target protein and the activation domain (AD) of the same transcription factor is linked to a peptide. If the protein and peptide interact BD and AD are brought together and the transcription factor reconstituted. This activates the transcription of a reporter gene

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