부산대학교 도서관

Streptavidin‐mediated enrichment is a powerful strategy to identify biotinylated biomolecules and their interaction partners; however, intense streptavidin‐derived peptides impede protein identification by mass spectrometry. Here, we present an approach to chemically modify streptavidin, thus rendering it resistant to proteolysis by trypsin and LysC. This modification results in over 100‐fold reduction of streptavidin contamination and in better coverage of proteins interacting with various biotinylated bait molecules (DNA, protein, and lipid) in an overall simplified workflow. Synopsis: Many proteomic studies rely on streptavidin‐based purifications. To avoid streptavidin contamination, this study presents a straightforward protocol to prevent its proteolytic digestion. Protein identification rates are improved in various applications. Streptavidin is rendered resistant to Trypsin/LysC digestion in two chemical reactions that modify argininyl and lysinyl residues.Protease‐resistant beads reduce contamination with streptavidin 100–1,000‐fold.Reduced contamination allows complete injection of samples without affecting LC‐MS performance.Reduced contamination improves protein sampling depth in interaction studies using biotin‐streptavidin enrichment, such as BioID and ChIP‐SICAP. [ABSTRACT FROM AUTHOR]