Authors:
Rui M M Branca, Lukas M Orre, Henrik J Johansson, Viktor Granholm, Mikael Huss, Åsa Pérez-Bercoff, Jenny Forshed, Lukas Käll, & Janne Lehtiö
Summary:
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.
Source:
Nature Methods; (11/17/13)