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Nat Biotechnol. 2002 May;20(5):512-5.

Quantitative proteome analysis by solid-phase isotope tagging and mass spectrometry.

Author information

1
Institute for Systems Biology, 1441 North 34th Street, Seattle, WA 98103-8904, USA.

Abstract

The adaptation of sequences of chemical reactions to a solid-phase format has been essential to the automation, reproducibility, and efficiency of a number of biotechnological processes including peptide and oligonucleotide synthesis and sequencing. Here we describe a method for the site-specific, stable isotopic labeling of cysteinyl peptides in complex peptide mixtures through a solid-phase capture and release process, and the concomitant isolation of the labeled peptides. The recovered peptides were analyzed by microcapillary liquid chromatography and tandem mass spectrometry (microLC-MS/MS) to determine their sequences and relative quantities. The method was used to detect galactose-induced changes in protein abundance in the yeast Saccharomyces cerevisiae. A side-by-side comparison with the isotope-coded affinity tag (ICAT) method demonstrated that the solid-phase method for stable isotope tagging of peptides is comparatively simpler, more efficient, and more sensitive.

PMID:
11981568
DOI:
10.1038/nbt0502-512
[Indexed for MEDLINE]

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