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

Authors
  • Mao, Xiaoyi1
  • Sokpor, Godwin2
  • Staiger, Jochen1
  • Nguyen, Huu Phuc2
  • Tuoc, Tran2
  • 1 University Medical Center Goettingen, 37077 Goettingen, Germany. , (Germany)
  • 2 Ruhr University of Bochum, 44791 Bochum, Germany. , (Germany)
Type
Published Article
Journal
STAR protocols
Publication Date
Jun 18, 2021
Volume
2
Issue
2
Pages
100503–100503
Identifiers
DOI: 10.1016/j.xpro.2021.100503
PMID: 33997817
Source
Medline
Keywords
Language
English
License
Unknown

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). © 2021 The Author(s).

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