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Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes

The aim of this study was to generate an antibody specific to Ki-67 acetylated at lysine 3180, whose existence was reported in an acetylome study (Scholz, C., B.T. Weinert, S.A. Wagner, P. Beli, Y. Miyake, J. Qi, L.J. Jensen, W. Streicher, A.R. McCarthy, N.J. Westwood, S. Lain, J. Cox, P. Matthias,...

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Detalles Bibliográficos
Autor principal: Takagi, Masatoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261705/
https://www.ncbi.nlm.nih.gov/pubmed/32490211
http://dx.doi.org/10.1016/j.bbrep.2019.100720
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author Takagi, Masatoshi
author_facet Takagi, Masatoshi
author_sort Takagi, Masatoshi
collection PubMed
description The aim of this study was to generate an antibody specific to Ki-67 acetylated at lysine 3180, whose existence was reported in an acetylome study (Scholz, C., B.T. Weinert, S.A. Wagner, P. Beli, Y. Miyake, J. Qi, L.J. Jensen, W. Streicher, A.R. McCarthy, N.J. Westwood, S. Lain, J. Cox, P. Matthias, M. Mann, J.E. Bradner, and C. Choudhary. 2015). Rabbits were immunized with a synthetic acetylated peptide corresponding to acetylated lysine 3180 of Ki-67 and the residues flanking it. The obtained antibody, referred to as Ab3180 in this study, was affinity purified with the antigen peptide and characterized. Immunoblot analysis of cell extracts using Ab3180 revealed that this antibody unexpectedly recognized a set of acetylated proteins unrelated to Ki-67. Ab3180-recognizable proteins were immunoprecipitated from cell extracts in a stringent condition and identified by mass-spec analysis as subunits of BAF (mammalian SWI/SNF) chromatin remodeling complexes. The unique specificity of Ab3180 will allow this antibody to be a useful tool for analyzing the acetylation of BAF complexes and its significance to the formation/function of BAF complexes.
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spelling pubmed-72617052020-06-01 Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes Takagi, Masatoshi Biochem Biophys Rep Article The aim of this study was to generate an antibody specific to Ki-67 acetylated at lysine 3180, whose existence was reported in an acetylome study (Scholz, C., B.T. Weinert, S.A. Wagner, P. Beli, Y. Miyake, J. Qi, L.J. Jensen, W. Streicher, A.R. McCarthy, N.J. Westwood, S. Lain, J. Cox, P. Matthias, M. Mann, J.E. Bradner, and C. Choudhary. 2015). Rabbits were immunized with a synthetic acetylated peptide corresponding to acetylated lysine 3180 of Ki-67 and the residues flanking it. The obtained antibody, referred to as Ab3180 in this study, was affinity purified with the antigen peptide and characterized. Immunoblot analysis of cell extracts using Ab3180 revealed that this antibody unexpectedly recognized a set of acetylated proteins unrelated to Ki-67. Ab3180-recognizable proteins were immunoprecipitated from cell extracts in a stringent condition and identified by mass-spec analysis as subunits of BAF (mammalian SWI/SNF) chromatin remodeling complexes. The unique specificity of Ab3180 will allow this antibody to be a useful tool for analyzing the acetylation of BAF complexes and its significance to the formation/function of BAF complexes. Elsevier 2019-12-14 /pmc/articles/PMC7261705/ /pubmed/32490211 http://dx.doi.org/10.1016/j.bbrep.2019.100720 Text en © 2019 The Author http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Takagi, Masatoshi
Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title_full Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title_fullStr Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title_full_unstemmed Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title_short Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes
title_sort generation of an antibody recognizing a set of acetylated proteins, including subunits of baf complexes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261705/
https://www.ncbi.nlm.nih.gov/pubmed/32490211
http://dx.doi.org/10.1016/j.bbrep.2019.100720
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