Mapping of domain-mediated protein-protein interaction by SPOT peptide assay

通过 SPOT 肽分析绘制结构域介导的蛋白质-蛋白质相互作用

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作者:Xiaoyi Mao, Godwin Sokpor, Jochen Staiger, Huu Phuc Nguyen, Tran Tuoc

Abstract

Identification of peptides mediating protein-protein interaction (PPI) is crucial for understanding the function of interlinked proteins in cellular processes and amino acid-associated diseases. Traditional PPI assays are laborious, involving the generation of many truncated proteins. SPOT peptide assay allows high-throughput detection of domains essential for PPI by synthesizing several hundred peptides on a cellulose membrane. Here, we present a rapid SPOT peptide protocol for identifying the binding motifs, which mediate interaction between the chromatin remodeling factors BAF155/BAF170 and the epigenetic factor Kdm6b. For complete details on the use and execution of this protocol, please refer to Narayanan et al. (2015).

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