TitleTargeted detection and quantitation of histone modifications from 1,000 cells.
Publication TypeJournal Article
Year of Publication2020
AuthorsAbshiru, Nebiyu A., Sikora Jacek W., Camarillo Jeannie M., Morris Juliette A., Compton Philip D., Lee Tak, Neelamraju Yaseswini, Haddox Samuel, Sheridan Caroline, Carroll Martin, Cripe Larry D., Tallman Martin S., Paietta Elisabeth M., Melnick Ari M., Thomas Paul M., Garrett-Bakelman Francine E., and Kelleher Neil L.
JournalPLoS One
Volume15
Issue10
Paginatione0240829
Date Published2020
ISSN1932-6203
KeywordsAcetylation, Cell Line, Chromatography, Liquid, Histones, Humans, Methylation, Peptides, Protein Processing, Post-Translational, Proteomics, Tandem Mass Spectrometry
Abstract

<p>Histone post-translational modifications (PTMs) create a powerful regulatory mechanism for maintaining chromosomal integrity in cells. Histone acetylation and methylation, the most widely studied histone PTMs, act in concert with chromatin-associated proteins to control access to genetic information during transcription. Alterations in cellular histone PTMs have been linked to disease states and have crucial biomarker and therapeutic potential. Traditional bottom-up mass spectrometry of histones requires large numbers of cells, typically one million or more. However, for some cell subtype-specific studies, it is difficult or impossible to obtain such large numbers of cells and quantification of rare histone PTMs is often unachievable. An established targeted LC-MS/MS method was used to quantify the abundance of histone PTMs from cell lines and primary human specimens. Sample preparation was modified by omitting nuclear isolation and reducing the rounds of histone derivatization to improve detection of histone peptides down to 1,000 cells. In the current study, we developed and validated a quantitative LC-MS/MS approach tailored for a targeted histone assay of 75 histone peptides with as few as 10,000 cells. Furthermore, we were able to detect and quantify 61 histone peptides from just 1,000 primary human stem cells. Detection of 37 histone peptides was possible from 1,000 acute myeloid leukemia patient cells. We anticipate that this revised method can be used in many applications where achieving large cell numbers is challenging, including rare human cell populations.</p>

DOI10.1371/journal.pone.0240829
Alternate JournalPLoS One
PubMed ID33104722
PubMed Central IDPMC7588077
Grant ListUG1 CA189859 / CA / NCI NIH HHS / United States
P30 CA044579 / CA / NCI NIH HHS / United States
P41 GM108569 / GM / NIGMS NIH HHS / United States
P30 CA008748 / CA / NCI NIH HHS / United States
P30 CA060553 / CA / NCI NIH HHS / United States
U10 CA180820 / CA / NCI NIH HHS / United States
U10 CA180827 / CA / NCI NIH HHS / United States
UG1 CA233290 / CA / NCI NIH HHS / United States
UG1 CA233270 / CA / NCI NIH HHS / United States
K08 CA169055 / CA / NCI NIH HHS / United States