Precursor ion scans for the targeted detection of stable-isotope-labeled peptides.

Details

Serval ID
serval:BIB_3FD8777941B0
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Precursor ion scans for the targeted detection of stable-isotope-labeled peptides.
Journal
Rapid communications in mass spectrometry
Author(s)
Colzani M., Bienvenut W.V., Faes E., Quadroni M.
ISSN
1097-0231[electronic]
Publication state
Published
Issued date
2009
Peer-reviewed
Oui
Volume
23
Number
22
Pages
3570-3578
Language
english
Notes
Publication types: Evaluation Studies ; Journal Article
Publication Status: ppublish
Abstract
Stable isotope labels are routinely introduced into proteomes for quantification purposes. Full labeling of cells in varying biological states, followed by sample mixing, fractionation and intensive data acquisition, is used to obtain accurate large-scale quantification of total protein levels. However, biological processes often affect only a small group of proteins for a short time, resulting in changes that are difficult to detect against the total proteome background. An alternative approach could be the targeted analysis of the proteins synthesized in response to a given biological stimulus. Such proteins can be pulse-labeled with a stable isotope by metabolic incorporation of 'heavy' amino acids. In this study we investigated the specific detection and identification of labeled proteins using acquisition methods based on Precursor Ion Scans (PIS) on a triple-quadrupole ion trap mass spectrometer. PIS-based methods were set to detect unique immonium ions originating from labeled peptides. Different labels and methods were tested in standard mixtures to optimize performance. We showed that, in comparison with an untargeted analysis on the same instrument, the approach allowed a several-fold increase in the specificity of detection of labeled proteins over unlabeled ones. The technique was applied to the identification of proteins secreted by human cells into growth media containing bovine serum proteins, allowing the preferential detection of labeled cellular proteins over unlabeled bovine ones. However, compared with untargeted acquisitions on two different instruments, the PIS-based strategy showed some limitations in sensitivity. We discuss possible perspectives of the technique.
Keywords
Cell Line, Tumor, Humans, Ions/chemistry, Isotope Labeling, Isotopes/chemistry, Isotopes/metabolism, Mass Spectrometry/methods, Melanoma/chemistry, Melanoma/metabolism, Peptides/chemistry, Proteins/chemistry, Proteins/metabolism, Proteomics/methods
Pubmed
Web of science
Create date
05/01/2010 14:19
Last modification date
20/08/2019 13:37
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