HiRIEF LC-MSMS enables deep proteome coverage and unbiased proteogenomics

作者：Branca Rui M M; Orre Lukas M; Johansson Henrik J; Granholm Viktor; Huss Mikael; Perez Bercoff Asa; Forshed Jenny; Kall Lukas; Lehtio Janne^*

来源：Nature Methods, 2014, 11(1): 59-+.

DOI：10.1038/NMETH.2732

We present a liquid chromatography-mass spectrometry (LC-MSMS)-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.

• 出版日期2014-1