HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics.

Branca, Rui M M; Orre, Lukas M; Johansson, Henrik J; Granholm, Viktor; Huss, Mikael; Pérez-Bercoff, Åsa; Forshed, Jenny; Käll, Lukas et al. · Nat Methods · 2014

basic_science · Level V

Where this comes from

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