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

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 pe...

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Autores principales: Udeshi, Namrata D, Pedram, Kayvon, Svinkina, Tanya, Fereshetian, Shaunt, Myers, Samuel A, Aygun, Ozan, Krug, Karsten, Clauser, Karl, Ryan, Dominic, Ast, Tslil, Mootha, Vamsi K, Ting, Alice Y, Carr, Steven A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2017
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8954634/
https://www.ncbi.nlm.nih.gov/pubmed/29039416
http://dx.doi.org/10.1038/nmeth.4465
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author Udeshi, Namrata D
Pedram, Kayvon
Svinkina, Tanya
Fereshetian, Shaunt
Myers, Samuel A
Aygun, Ozan
Krug, Karsten
Clauser, Karl
Ryan, Dominic
Ast, Tslil
Mootha, Vamsi K
Ting, Alice Y
Carr, Steven A
author_facet Udeshi, Namrata D
Pedram, Kayvon
Svinkina, Tanya
Fereshetian, Shaunt
Myers, Samuel A
Aygun, Ozan
Krug, Karsten
Clauser, Karl
Ryan, Dominic
Ast, Tslil
Mootha, Vamsi K
Ting, Alice Y
Carr, Steven A
author_sort Udeshi, Namrata D
collection PubMed
description 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.
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spelling pubmed-89546342022-03-25 Antibodies to biotin enable large-scale detection of biotinylation sites on proteins Udeshi, Namrata D Pedram, Kayvon Svinkina, Tanya Fereshetian, Shaunt Myers, Samuel A Aygun, Ozan Krug, Karsten Clauser, Karl Ryan, Dominic Ast, Tslil Mootha, Vamsi K Ting, Alice Y Carr, Steven A Nat Methods Article 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. 2017-12 2017-10-16 /pmc/articles/PMC8954634/ /pubmed/29039416 http://dx.doi.org/10.1038/nmeth.4465 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use. Reprints and permissions information is available online at http://www.nature.com/reprints/index.html.
spellingShingle Article
Udeshi, Namrata D
Pedram, Kayvon
Svinkina, Tanya
Fereshetian, Shaunt
Myers, Samuel A
Aygun, Ozan
Krug, Karsten
Clauser, Karl
Ryan, Dominic
Ast, Tslil
Mootha, Vamsi K
Ting, Alice Y
Carr, Steven A
Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title_full Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title_fullStr Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title_full_unstemmed Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title_short Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
title_sort antibodies to biotin enable large-scale detection of biotinylation sites on proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8954634/
https://www.ncbi.nlm.nih.gov/pubmed/29039416
http://dx.doi.org/10.1038/nmeth.4465
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