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

TitleMARQUIS: a multiplex method for absolute quantification of peptides and posttranslational modifications.
Publication TypeJournal Article
Year of Publication2015
AuthorsCurran, TG, Zhang, Y, Ma, DJ, Sarkaria, JN, White, FM
JournalNat Commun
Volume6
Pagination5924
Date Published2015
ISSN2041-1723
KeywordsAnimals, Cell Line, Tumor, Computational Biology, Glioblastoma, Humans, Ligands, Mass Spectrometry, Mice, Neoplasm Transplantation, Peptides, Phosphorylation, Protein Binding, Protein Processing, Post-Translational, Proteomics, Receptor, Epidermal Growth Factor, Signal Transduction, Tyrosine
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.

DOI10.1038/ncomms6924
Alternate JournalNat Commun
PubMed ID25581283
PubMed Central IDPMC4293043
Grant ListCA108961 / CA / NCI NIH HHS / United States
P50 CA108961 / CA / NCI NIH HHS / United States
R01 CA096504 / CA / NCI NIH HHS / United States
R01 CA096504 / CA / NCI NIH HHS / United States
R01 CA118705 / CA / NCI NIH HHS / United States
R01 CA118705 / CA / NCI NIH HHS / United States
U01 CA154966 / CA / NCI NIH HHS / United States
U54 CA112967 / CA / NCI NIH HHS / United States
U54 CA112967 / CA / NCI NIH HHS / United States