HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics
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Branca, Rui M M
Orre, Lukas M
Johansson, Henrik J
Granholm, Viktor
Huss, Mikael
Perez-Bercoff, Asa
Forshed, Jenny
Käll, Lukas
Lehtiö, Janne
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Nature Publishing Group
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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.
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Nature Methods: techniques for life scientists and chemists
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2037-12-31