MassSQUIRM: An assay for quantitative measurement of lysine demethylase activity

Lauren P. Blair, Nathan L. Avaritt, Rong Huang, Phillip A. Cole, Sean Dixon Taverna, Alan J. Tackett

Research output: Contribution to journalArticle

Abstract

In eukaryotes, DNA is wrapped around proteins called histones and is condensed into chromatin. Post-translational modification of histones can result in changes in gene expression. One of the most well-studied histone modifications is the methylation of lysine 4 on histone H3 (H3K4). This residue can be mono-, di- or tri-methylated and these varying methylation states have been associated with different levels of gene expression. Understanding exactly what the purpose of these methylation states is, in terms of gene expression, has been a topic of much research in recent years. Enzymes that can add (methyltransferases) and remove (demethylases) these modifications are of particular interest. The first demethylase discovered, LSD1, is the most well-classified and has been implicated in contributing to human cancers and to DNA damage response pathways. Currently, there are limited methods for accurately studying the activity of demethylases in vitro or in vivo. In this work, we present MassSQUIRM (mass spectrometric quantitation using isotopic reductive methylation), a quantitative method for studying the activity of demethylases capable of removing mono- and di-methyl marks from lysine residues. We focus specifically on LSD1 due to its potential as a prime therapeutic target for human disease. This quantitative approach will enable better characterization of the activity of LSD1 and other chromatin modifying enzymes in vitro, in vivo or in response to inhibitors.

Original languageEnglish (US)
Pages (from-to)491-500
Number of pages10
JournalEpigenetics
Volume6
Issue number4
DOIs
StatePublished - Apr 2011

Fingerprint

Methylation
Lysine
Histones
Gene Expression
Chromatin
Histone Code
Methyltransferases
Enzymes
Post Translational Protein Processing
Eukaryota
DNA Damage
DNA
Research
Neoplasms
Proteins
In Vitro Techniques
Therapeutics

Keywords

  • LSD1
  • Lysine demethylase
  • Mass spectrometry
  • Monoamine oxidase (MAO) inhibitors
  • Reductive methylation

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

Cite this

Blair, L. P., Avaritt, N. L., Huang, R., Cole, P. A., Taverna, S. D., & Tackett, A. J. (2011). MassSQUIRM: An assay for quantitative measurement of lysine demethylase activity. Epigenetics, 6(4), 491-500. https://doi.org/10.4161/epi.6.4.14531

MassSQUIRM : An assay for quantitative measurement of lysine demethylase activity. / Blair, Lauren P.; Avaritt, Nathan L.; Huang, Rong; Cole, Phillip A.; Taverna, Sean Dixon; Tackett, Alan J.

In: Epigenetics, Vol. 6, No. 4, 04.2011, p. 491-500.

Research output: Contribution to journalArticle

Blair, LP, Avaritt, NL, Huang, R, Cole, PA, Taverna, SD & Tackett, AJ 2011, 'MassSQUIRM: An assay for quantitative measurement of lysine demethylase activity', Epigenetics, vol. 6, no. 4, pp. 491-500. https://doi.org/10.4161/epi.6.4.14531
Blair, Lauren P. ; Avaritt, Nathan L. ; Huang, Rong ; Cole, Phillip A. ; Taverna, Sean Dixon ; Tackett, Alan J. / MassSQUIRM : An assay for quantitative measurement of lysine demethylase activity. In: Epigenetics. 2011 ; Vol. 6, No. 4. pp. 491-500.
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