Dataset Information

A mass-tolerant database search identifies a large proportion of unassigned spectra in shotgun proteomics as modified peptides

ABSTRACT: Less than half of all MS/MS spectra acquired in shotgun proteomics typically result in a confident peptide match. Here we present an ultra-tolerant Sequest database search that allowed peptide matching even with modifications of unknown masses up to ±500 Da. From an HEK293 cell proteome-wide dataset (9,513 proteins and 396,736 peptides), a ±500-Da search matched an additional 184,000 modified peptides. These were linked to both biological and chemical modifications representing 523 distinct mass bins including phosphorylation, glycosylation, and methylation. We attempted to localize all unknown modification masses to specific regions within a peptide, and known modifications were accurately assigned to the correct amino acids with frequencies often >90%. These data demonstrate that a large fraction of previously unassignable spectra are assignable to peptide sequences with modifications.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain, Kidney

SUBMITTER: Joel Chick

LAB HEAD: Steven P Gygi

PROVIDER: PXD001468 | Pride | 2015-06-03

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	SupplementaryTable1.xlsb	Xls
	SupplementaryTable2.xlsb	Xls
	SupplementaryTable3.xlsx	Xlsx
	b1906_293T_proteinID_01A_QE3_122212.mzXML	Mzxml
	b1906_293T_proteinID_01A_QE3_122212.raw	Raw

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A mass-tolerant database search identifies a large proportion of unassigned spectra in shotgun proteomics as modified peptides.

Chick Joel M JM Kolippakkam Deepak D Nusinow David P DP Zhai Bo B Rad Ramin R Huttlin Edward L EL Gygi Steven P SP

Nature biotechnology 20150615 7

Fewer than half of all tandem mass spectrometry (MS/MS) spectra acquired in shotgun proteomics experiments are typically matched to a peptide with high confidence. Here we determine the identity of unassigned peptides using an ultra-tolerant Sequest database search that allows peptide matching even with modifications of unknown masses up to ± 500 Da. In a proteome-wide data set on HEK293 cells (9,513 proteins and 396,736 peptides), this approach matched an additional 184,000 modified peptides, w ...[more]

PMID: 26076430

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