Proteomics

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A Collision-induced Dissociation Cleavable Isobaric Tag for Peptide Fragment Ion-based Quantification in Proteomics


ABSTRACT: Fragment ion-based proteome quantification methods quantify peptides based on unique peptide fragment ions avoiding the issue of ratio distortion that is commonly observed for reporter ion-based quantification approaches. Herein, we present a novel approach that relies on a collision-induced dissociation (CID)-cleavable isobaric acetyl-isoleucine-proline-glycine (Ac-IPG) tag, which conserves the merits of quantifying peptides based on unique fragment ions while reducing the complexity of the b-ion series thus facilitating data processing. Multiplex labelling is based on selective N-terminal dimethylation followed by derivatizing the C-terminal Lys residue of LysC peptides with isobaric Ac-IPG tags with different isotope distributions on Pro-Gly and Ac-Ile. Upon fragmentation between Ile and Pro, the resulting y-ions with the neutral loss of Ac-Ile can be distinguished between the different labelling channels based on different numbers of isotope labels on the Pro-Gly part. The peak intensities of y-ions contain the information for relative quantification.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Xiaobo Tian  

LAB HEAD: Rainer Bischoff

PROVIDER: PXD018790 | Pride | 2020-08-17

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
3-plexAc-IPGBSA1510yeast125April23.mzid Mzid
3-plexAc-IPGyeast111April232020.mzid Mzid
Ac-IPG-BSA1510_yeast_125.mgf Mgf
Ac-IPG-BSA1510_yeast_125.raw Raw
Ac-IPG-yeast-1-1-1.mgf Mgf
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Publications

A Collision-Induced Dissociation Cleavable Isobaric Tag for Peptide Fragment Ion-Based Quantification in Proteomics.

Tian Xiaobo X   de Vries Marcel P MP   Permentier Hjalmar P HP   Bischoff Rainer R  

Journal of proteome research 20200817 9


Quantifying peptides based on unique peptide fragment ions avoids the issue of ratio distortion that is commonly observed for reporter ion-based quantification approaches. Herein, we present a collision-induced dissociation-cleavable, isobaric acetyl-isoleucine-proline-glycine (Ac-IPG) tag, which conserves the merits of quantifying peptides based on unique fragments while reducing the complexity of the b-ion series compared to conventional fragment ion-based quantification methods thus facilitat  ...[more]

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