I am planning to analyze post-translational modifications, including acetylation and methylation of human histones, H3 and H4, via mass spectrometry (LC-ESI-Q-ToF). Can I ask for some advice and recommended protocols of histones isolation?
Dear Emilia:
I would check Methods in Enzymology.
Best of luck.
1. Methods Enzymol. 2012;512:3-28. doi: 10.1016/B978-0-12-391940-3.00001-9.
Examining histone posttranslational modification patterns by high-resolution mass spectrometry.
Free full text article:
Lin S, Garcia BA.
Histone variants and posttranslational modifications (PTMs) are essential for epigenetic regulation of transcriptional expression. Single and/or combinatorial PTMs of histones play important roles in development and disease formation. Mass spectrometry (MS) has been a powerful tool to study histone variants and PTMs, as it not only can identify novel PTMs but also can provide quantitative measurement of a spectrum of histone variants and PTMs in the same sample. In this chapter, we employ a combination of chemical derivation and high-resolution MS to identify and quantify multiple histone variants and PTMs. Histones are acid extracted and modified with propionyl groups and subsequently produces suitable sizes of fragments for MS analysis by trypsin digestion. The newly generated N-termini of histone peptides can be differentially marked by stable isotope labeling in a second reaction of propionylation, which enables direct comparison between two different samples in the following MS analysis.
2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC223280/pdf/pnas00114-0128.pdf
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