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Antibodies to biotin enable large-scale detection of biotinylation sites on proteins.

Namrata D UdeshiKayvon PedramTanya SvinkinaShaunt FereshetianSamuel A MyersOzan AygunKarsten KrugKarl ClauserDominic RyanTslil AstVamsi K MoothaAlice Y TingSteven A Carr
Published in: Nature methods (2017)
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.
Keyphrases
  • highly efficient
  • mass spectrometry
  • liquid chromatography
  • hydrogen peroxide
  • high resolution
  • nitric oxide
  • real time pcr
  • label free
  • sensitive detection
  • simultaneous determination