A novel strategy for quantitative proteomics using isotope‐coded protein labels
Top Cited Papers
- 25 January 2005
- journal article
- research article
- Published by Wiley in Proteomics
- Vol. 5 (1) , 4-15
- https://doi.org/10.1002/pmic.200400873
Abstract
Stable isotope labelling in combination with mass spectrometry has emerged as a powerful tool to identify and relatively quantify thousands of proteins within complex protein mixtures. Here we describe a novel method, termed isotope‐coded protein label (ICPL), which is capable of high‐throughput quantitative proteome profiling on a global scale. Since ICPL is based on stable isotope tagging at the frequent free amino groups of isolated intact proteins, it is applicable to any protein sample, including extracts from tissues or body fluids, and compatible to all separation methods currently employed in proteome studies. The method showed highly accurate and reproducible quantification of proteins and yielded high sequence coverage, indispensable for the detection of post‐translational modifications and protein isoforms. The efficiency (e.g. accuracy, dynamic range, sensitivity, speed) of the approach is demonstrated by comparative analysis of two differentially spiked proteomes.Keywords
This publication has 38 references indexed in Scilit:
- Approaches for the quantification of protein concentration ratiosProteomics, 2003
- Metabolic labeling of C. elegans and D. melanogaster for quantitative proteomicsNature Biotechnology, 2003
- Two-dimensional gel electrophoresis; better than a poke in the ICAT?Current Opinion in Biotechnology, 2002
- Controlling Deuterium Isotope Effects in Comparative ProteomicsAnalytical Chemistry, 2002
- Web-based two-dimensional database ofSaccharomyces cerevisiaeproteins using immobilized pH gradients from pH 6 to pH 12 and matrix-assisted laser desorption/ionization-time of flight mass spectrometryProteomics, 2002
- Genetic analysis of the mouse brain proteomeNature Genetics, 2002
- Preparative two-dimensional gel electrophoresis at alkaline pH using narrow range immobilized pH gradientsProteomics, 2002
- Large-scale analysis of the yeast proteome by multidimensional protein identification technologyNature Biotechnology, 2001
- Quantitative analysis of complex protein mixtures using isotope-coded affinity tagsNature Biotechnology, 1999
- Method to Correlate Tandem Mass Spectra of Modified Peptides to Amino Acid Sequences in the Protein DatabaseAnalytical Chemistry, 1995