HiRIEF LC-MS enables deep proteome coverage and unbiased proteogenomics.
Rui M M BrancaLukas M OrreHenrik J JohanssonViktor GranholmMikael HussÅsa Pérez-BercoffJenny ForshedLukas KällJanne LehtiöPublished in: Nature methods (2013)
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.
Keyphrases
- genome wide
- mass spectrometry
- high resolution
- liquid chromatography
- endothelial cells
- dna methylation
- copy number
- induced pluripotent stem cells
- pluripotent stem cells
- tandem mass spectrometry
- high resolution mass spectrometry
- small molecule
- gene expression
- genome wide association study
- high performance liquid chromatography
- simultaneous determination
- working memory
- binding protein
- healthcare
- capillary electrophoresis
- molecular dynamics simulations