MARQUIS: A multiplex method for absolute quantification of peptides and posttranslational modifications

Timothy G. Curran, Yi Zhang, Daniel J. Ma, Jann N Sarkaria, Forest M. White

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Absolute quantification of protein expression and posttranslational modifications by mass spectrometry has been challenging due to a variety of factors, including the potentially large dynamic range of phosphorylation response. To address these issues, we have developed MARQUIS-Multiplex Absolute Regressed Quantification with Internal Standards-a novel mass spectrometry-based approach using a combination of isobaric tags and heavy-labelled standard peptides, to construct internal standard curves for peptides derived from key nodes in signal transduction networks. We applied MARQUIS to quantify phosphorylation dynamics within the EGFR network at multiple time points following stimulation with several ligands, enabling a quantitative comparison of EGFR phosphorylation sites and demonstrating that receptor phosphorylation is qualitatively similar but quantitatively distinct for each EGFR ligand tested. MARQUIS was also applied to quantify the effect of EGFR kinase inhibition on glioblastoma patient-derived xenografts. MARQUIS is a versatile method, broadly applicable and extendable to multiple mass spectrometric platforms.

Original languageEnglish (US)
Article number5924
JournalNature Communications
Volume6
DOIs
StatePublished - 2015

Fingerprint

phosphorylation
Phosphorylation
Post Translational Protein Processing
peptides
Peptides
Mass spectrometry
Mass Spectrometry
mass spectroscopy
Ligands
Signal transduction
ligands
Glioblastoma
stimulation
Heterografts
dynamic range
Signal Transduction
Phosphotransferases
platforms
proteins
curves

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

MARQUIS : A multiplex method for absolute quantification of peptides and posttranslational modifications. / Curran, Timothy G.; Zhang, Yi; Ma, Daniel J.; Sarkaria, Jann N; White, Forest M.

In: Nature Communications, Vol. 6, 5924, 2015.

Research output: Contribution to journalArticle

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