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01/12/17 |
Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
Nature Methods. Jan-12-2017;14(12):1167 - 1170. doi: 10.1038/nmeth.4465
Although purification of biotinylated molecules is highly efficient, identifying specific sites of biotinylation remains challenging. We show that anti-biotin antibodies enable unprecedented enrichment of biotinylated peptides from complex peptide mixtures. Live-cell proximity labeling using APEX peroxidase followed by anti-biotin enrichment and mass spectrometry yielded over 1,600 biotinylation sites on hundreds of proteins, an increase of more than 30-fold in the number of biotinylation sites identified compared to streptavidin-based enrichment of proteins.