A Rapid and Efficient Method for the Extraction of Histone Proteins.
J Proteome Res
; 22(8): 2765-2773, 2023 08 04.
Article
em En
| MEDLINE
| ID: mdl-37463329
ABSTRACT
Current protocols used to extract and purify histones are notoriously tedious, especially when using yeast cells. Here, we describe the use of a simple filter-aided sample preparation approach enabling histone extraction from yeast and mammalian cells using acidified ethanol, which not only improves extraction but also inactivates histone-modifying enzymes. We show that our improved method prevents N-terminal clipping of H3, an artifact frequently observed in yeast cells using standard histone extraction protocols. Our method is scalable and provides efficient recovery of histones when extracts are prepared from as few as two million yeast cells. We further demonstrate the application of this approach for the analysis of histone modifications in fungal clinical isolates available in a limited quantity. Compared with standard protocols, our method enables the study of histones and their modifications in a faster, simpler, and more robust manner.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
/
Histonas
Tipo de estudo:
Guideline
Limite:
Animals
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article