HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics.
basic_science · Level V
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- Record sourced from PubMed, PMID 24240322.
- Also identified by DOI 10.1038/nmeth.2732.
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Abstract
We present a liquid chromatography-mass spectrometry (LC-MS)-based method permitting unbiased (gene prediction-independent) genome-wide discovery of protein-coding loci in higher eukaryotes. Using high-resolution isoelectric focusing (HiRIEF) at the peptide level in the 3.7-5.0 pH range and accurate peptide isoelectric point (pI) prediction, we probed the six-reading-frame translation of the human and mouse genomes and identified 98 and 52 previously undiscovered protein-coding loci, respectively. The method also enabled deep proteome coverage, identifying 13,078 human and 10,637 mouse proteins.
Medical subject headings
- Chromatography, Liquid
- Genomics
- Mass Spectrometry
- Proteome
- Proteomics