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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.
Afiliação
  • Udeshi ND; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Pedram K; Departments of Genetics, Biology, and Chemistry, Stanford University, Stanford, California, USA.
  • Svinkina T; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Fereshetian S; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Myers SA; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Aygun O; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Krug K; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Clauser K; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Ryan D; DRI2 LLC, Littleton, Massachusetts, USA.
  • Ast T; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Mootha VK; Howard Hughes Medical Institute, Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, USA.
  • Ting AY; Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
  • Carr SA; Howard Hughes Medical Institute, Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, USA.
Nat Methods ; 14(12): 1167-1170, 2017 Dec.
Article em En | MEDLINE | ID: mdl-29039416
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Temas: Geral Base de dados: MEDLINE Assunto principal: Peptídeos / Biotina / Proteínas / Anticorpos Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Revista: Nat Methods Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Temas: Geral Base de dados: MEDLINE Assunto principal: Peptídeos / Biotina / Proteínas / Anticorpos Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Revista: Nat Methods Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Estados Unidos