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. 2014 Jan;11(1):59-62.
doi: 10.1038/nmeth.2732. Epub 2013 Nov 17.

HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics

Rui M M Branca  1 , Lukas M Orre  1 , Henrik J Johansson  1 , Viktor Granholm  2 , Mikael Huss  3 , Åsa Pérez-Bercoff  4 , Jenny Forshed  4 , Lukas Käll  5 , Janne Lehtiö  4
Affiliations

HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics

Rui M M Branca et al. Nat Methods. 2014 Jan.

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.

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