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diDO-IPTL: A Peptide-Labeling Strategy for Precision Quantitative Proteomics
- Waldbauer, Jacob, Zhang, Lichun, Rizzo, Adriana, Muratore, Daniel
- Analytical chemistry 2017 v.89 no.21 pp. 11498-11504
- algorithms, formaldehyde, ions, oxygen, peptides, proteins, proteomics, stable isotopes
- We present an analytical strategy, dimethylation-deuteration and oxygen-exchange IPTL (diDO-IPTL), for high-precision, broad-coverage quantitative proteomics. The diDO-IPTL approach combines two advances in isobaric peptide terminal labeling (IPTL) methodology: first, a one-pot chemical labeling strategy for attaching isotopic tags to both the N- and C-termini of tryptic peptides, and second, a search engine (based on the Morpheus algorithm) optimized for identification and quantification of twinned peaks from peptide fragment ions in MS² spectra. The diDO-IPTL labeling chemistry uses only high-purity, relatively inexpensive isotopic reagents (¹⁸O water and deuterated formaldehyde) and requires no postlabeling cleanup or isotopic impurity corrections. This strategy produces proteome-scale relative quantification results with high accuracy and precision, suitable for the detection of small protein abundance variations between complex biological samples. In a two-proteome mixture experiment, diDO-IPTL quantification discriminates 1.5-fold changes in abundance of over 1000 proteins with 88% accuracy. The diDO-IPTL methodology is a high-precision, economical approach to quantitative proteomics that is applicable to a wide variety of sample types.