Dataset Information

DeMix workflow for efficient identification of cofragmented peptides in high resolution data-dependent tandem mass spectrometry.

ABSTRACT: Based on conventional data-dependent acquisition strategy of shotgun proteomics, we present a new workflow DeMix, which significantly increases the efficiency of peptide identification for in-depth shotgun analysis of complex proteomes. Capitalizing on the high resolution and mass accuracy of Orbitrap-based tandem mass spectrometry, we developed a simple deconvolution method of "cloning" chimeric tandem spectra for cofragmented peptides. Additional to a database search, a simple rescoring scheme utilizes mass accuracy and converts the unwanted cofragmenting events into a surprising advantage of multiplexing. With the combination of cloning and rescoring, we obtained on average nine peptide-spectrum matches per second on a Q-Exactive workbench, whereas the actual MS/MS acquisition rate was close to seven spectra per second. This efficiency boost to 1.24 identified peptides per MS/MS spectrum enabled analysis of over 5000 human proteins in single-dimensional LC-MS/MS shotgun experiments with an only two-hour gradient. These findings suggest a change in the dominant "one MS/MS spectrum - one peptide" paradigm for data acquisition and analysis in shotgun data-dependent proteomics. DeMix also demonstrated higher robustness than conventional approaches in terms of lower variation among the results of consecutive LC-MS/MS runs.

SUBMITTER: Zhang B

PROVIDER: S-EPMC4223503 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature

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DeMix workflow for efficient identification of cofragmented peptides in high resolution data-dependent tandem mass spectrometry.

Zhang Bo B Pirmoradian Mohammad M Chernobrovkin Alexey A Zubarev Roman A RA

Molecular & cellular proteomics : MCP 20140806 11

Based on conventional data-dependent acquisition strategy of shotgun proteomics, we present a new workflow DeMix, which significantly increases the efficiency of peptide identification for in-depth shotgun analysis of complex proteomes. Capitalizing on the high resolution and mass accuracy of Orbitrap-based tandem mass spectrometry, we developed a simple deconvolution method of "cloning" chimeric tandem spectra for cofragmented peptides. Additional to a database search, a simple rescoring scheme ...[more]

PMID: 25100859

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Dataset Information

DeMix workflow for efficient identification of cofragmented peptides in high resolution data-dependent tandem mass spectrometry.

Publications

DeMix workflow for efficient identification of cofragmented peptides in high resolution data-dependent tandem mass spectrometry.

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