Filter-Aided Sample Preparation Procedure for Mass Spectrometric Analysis of Plant Histones

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Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Středoevropský technologický institut. Oficiální stránka publikace je na webu muni.cz.
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LEDVINOVÁ Dominika MIKULÁŠEK Kamil KUCHAŘÍKOVÁ Hana BRABENCOVÁ Sylva FOJTOVÁ Miloslava ZDRÁHAL Zbyněk LOCHMANOVÁ Gabriela

Rok publikování 2018
Druh Článek v odborném periodiku
Časopis / Zdroj Frontiers in Plant Science
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
Doi http://dx.doi.org/10.3389/fpls.2018.01373
Klíčová slova histone derivatization; filter-aided sample preparation; post-translational modifications; epigenetics; mass spectrometry; Arabidopsis thaliana
Popis Characterization of histone post-translational modifications (PTMs) is still challenging, and robust histone sample preparation is essential for convincing evaluation of PTMs by mass spectrometry. An effective protocol for extracting plant histone proteins must also avoid excessive co-extraction of the numerous potential interfering compounds, including those related to secondary metabolism. Currently, the co-existence of histone marks is addressed mostly by shotgun proteomic analysis following chemical derivatization of histone lysine residues. Here, we report a straightforward approach for plant histone sample preparation for mass spectrometry, based on filter-aided sample preparation coupled with histone propionylation. The approach offers savings in sample handling and preparation time, enables removal of interfering compounds from the sample, and does not require either precipitation or dialysis of histone extract. We show the comparison of two protocol variants for derivatization of histone proteins, in-solution propionylation in the vial and propionylation on the filter unit. For both protocols, we obtained identical abundances of post-translationally modified histone peptides. Although shorter time is required for histone protein labeling on the filter unit, in-solution derivatization slightly outweighed filter-based variant by lower data variability. Nevertheless, both protocol variants appear to be efficient and convenient approach for preparation of plant histones for mass spectrometric analysis.
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