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

janelia7_blocks-janelia7_biblio_header | block

Nature Methods. Jan-12-2017;14(12):1167 - 1170. doi: 10.1038/nmeth.4465

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

Udeshi ND, Pedram K, Svinkina T, Fereshetian S, Myers SA, Aygun O, Krug K, Clauser K, Ryan D, Ast T, Mootha VK, Ting AY, Carr SA

Note: Research in this publication was not performed at Janelia.

janelia7_blocks-janelia7_biblio_abstract | block

Abstract

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.

node:body | entity_field

Main Menu (Mobile)- Block

Main Menu - Block

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

Note: Research in this publication was not performed at Janelia.

Abstract

Janelia Authors