High-throughput profiling of histone post-translational modifications and chromatin modifying proteins by reverse phase protein array.

Animals Chromatin Epigenesis, Genetic Histones / metabolism Mice Protein Array Analysis / methods Protein Processing, Post-Translational
Breast cancer Epigenetics High-throughput Induced pluripotent stem cells Post-translational modifications RPPA

Journal

Journal of proteomics
ISSN: 1876-7737
Titre abrégé: J Proteomics
Pays: Netherlands
ID NLM: 101475056

Informations de publication

Date de publication:
30 06 2022
Historique:
received: 28 07 2021
revised: 23 03 2022
accepted: 11 04 2022
pubmed: 1 5 2022
medline: 26 5 2022
entrez: 30 4 2022
Statut: ppublish

Résumé

Epigenetic variation plays a significant role in normal development and human diseases including cancer, in part through post-translational modifications (PTMs) of histones. Identification and profiling of changes in histone PTMs, and in proteins regulating PTMs, are crucial to understanding diseases, and for discovery of epigenetic therapeutic agents. In this study, we have adapted and validated an antibody-based reverse phase protein array (RPPA) platform for profiling 20 histone PTMs and expression of 40 proteins that modify histones and other epigenomic regulators. The specificity of the RPPA assay for histone PTMs was validated with synthetic peptides corresponding to histone PTMs and by detection of histone PTM changes in response to inhibitors of histone modifier proteins in cell cultures. The useful application of the RPPA platform was demonstrated with two models: induction of pluripotent stem cells and a mouse mammary tumor progression model. Described here is a robust platform that includes a rapid microscale method for histone isolation and partially automated workflows for analysis of histone PTMs and histone modifiers that can be performed in a high-throughput manner with hundreds of samples. This RPPA platform has potential for translational applications through the discovery and validation of epigenetic states as therapeutic targets and biomarkers. SIGNIFICANCE: Our study has established an antibody-based reverse phase protein array platform for global profiling of a wide range of post-translational modifications of histones and histone modifier proteins. The high-throughput platform provides comprehensive analyses of epigenetics for biological research and disease studies and may serve as screening assay for diagnostic purpose or therapy development.

Identifiants

DOI: 10.1016/j.jprot.2022.104596 PMID: 35489683 PMC: PMC10165948
pubmed: 35489683
pii: S1874-3919(22)00120-8
doi: 10.1016/j.jprot.2022.104596
pmc: PMC10165948
mid: NIHMS1892523
pii:
doi:

Substances chimiques

Chromatin 0
Histones 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

104596

Subventions

Organisme : NIA NIH HHS
ID : P01 AG066606
Pays : United States
Organisme : NIH HHS
ID : S10 OD028591
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA193235
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM139295
Pays : United States
Organisme : NIH HHS
ID : S10 OD028648
Pays : United States
Organisme : NIEHS NIH HHS
ID : P30 ES030285
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA125123
Pays : United States

Informations de copyright

Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.

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Auteurs

Xuan Wang (X)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA.

Zhongcheng Shi (Z)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA.

Hsin-Yi Lu (HY)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA.

Jean J Kim (JJ)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA.

Wen Bu (W)

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Lester & Sue Smith Breast Center, Baylor College of Medicine, Houston, TX, USA.

Jose A Villalobos (JA)

Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Dimuthu N Perera (DN)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA.

Sung Yun Jung (SY)

Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Tao Wang (T)

Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Sandra L Grimm (SL)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA.

Bethany C Taylor (BC)

Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Kimal Rajapakshe (K)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA.

Hyekyung Park (H)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA.

Julia Wulfkuhle (J)

Center for Applied Proteomics and Molecular Medicine, George Mason University, Fairfax, VA, USA.

Nicolas L Young (NL)

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA; Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Yi Li (Y)

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Lester & Sue Smith Breast Center, Baylor College of Medicine, Houston, TX, USA.

Cristian Coarfa (C)

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA.

Dean P Edwards (DP)

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA.

Shixia Huang (S)

Advanced Technology Cores, Baylor College of Medicine, Houston, TX, USA; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA; Department of Education, Innovation & Technology, Baylor College of Medicine, Houston, TX, USA. Electronic address: shixiah@bcm.edu.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › High-throughput profiling of histone...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Structures de chromosome › Chromatine › High-throughput profiling of histone...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › High-throughput profiling of histone...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Nucléoprotéines › Histone › High-throughput profiling of histone...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › High-throughput profiling of histone...
questionsmedicales.fr › Techniques d'investigation › Techniques de sonde moléculaire › Analyse par réseau de protéines › High-throughput profiling of histone...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Modification traductionnelle des protéines › Maturation post-traductionnelle des protéines › High-throughput profiling of histone...