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

@article{1442900,
   author = {Ledvinová, Dominika and Mikulášek, Kamil and Kuchaříková, Hana and Brabencová, Sylva and Fojtová, Miloslava and Zdráhal, Zbyněk and Lochmanová, Gabriela},
   article_number = {SEP},
   doi = {http://dx.doi.org/10.3389/fpls.2018.01373},
   keywords = {histone derivatization; filter-aided sample preparation; post-translational modifications; epigenetics; mass spectrometry; Arabidopsis thaliana},
   language = {eng},
   issn = {1664-462X},
   journal = {Frontiers in Plant Science},
   title = {Filter-Aided Sample Preparation Procedure for Mass Spectrometric Analysis of Plant Histones},
   volume = {9},
   year = {2018}
}

TY  - JOUR
ID  - 1442900
AU  - Ledvinová, Dominika - Mikulášek, Kamil - Kuchaříková, Hana - Brabencová, Sylva - Fojtová, Miloslava - Zdráhal, Zbyněk - Lochmanová, Gabriela
PY  - 2018
TI  - Filter-Aided Sample Preparation Procedure for Mass Spectrometric Analysis of Plant Histones
JF  - Frontiers in Plant Science
VL  - 9
IS  - SEP
SP  - 1373
EP  - 1373
PB  - Frontiers Research Foundation
SN  - 1664462X
KW  - histone derivatization
KW  - filter-aided sample preparation
KW  - post-translational modifications
KW  - epigenetics
KW  - mass spectrometry
KW  - Arabidopsis thaliana
N2  - 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.
ER  - 

LEDVINOVÁ, Dominika, Kamil MIKULÁŠEK, Hana KUCHAŘÍKOVÁ, Sylva BRABENCOVÁ, Miloslava FOJTOVÁ, Zbyněk ZDRÁHAL and Gabriela LOCHMANOVÁ. Filter-Aided Sample Preparation Procedure for Mass Spectrometric Analysis of Plant Histones. \textit{Frontiers in Plant Science}. Frontiers Research Foundation, 2018, vol.~9, SEP, p.~1373-1383. ISSN~1664-462X. Available from: https://dx.doi.org/10.3389/fpls.2018.01373.