DirectMS1: MS/MS-Free Identification of 1000 Proteins of Cellular Proteomes in 5 Minutes.
Mikhail V GorshkovJulia A BubisVladimir A GorshkovIrina A TarasovaLev I LevitskyAnna A LobasElizaveta M SolovyevaMarina L PridatchenkoFrank KjeldsenMikhail V GorshkovPublished in: Analytical chemistry (2020)
Proteome characterization relies heavily on tandem mass spectrometry (MS/MS) and is thus associated with instrumentation complexity, lengthy analysis time, and limited duty cycle. It was always tempting to implement approaches that do not require MS/MS, yet they were constantly failing to achieve a meaningful depth of quantitative proteome coverage within short experimental times, which is particularly important for clinical or biomarker-discovery applications. Here, we report on the first successful attempt to develop a truly MS/MS-free method, DirectMS1, for bottom-up proteomics. The method is compared with the standard MS/MS-based data-dependent acquisition approach for proteome-wide analysis using 5 min LC gradients. Specifically, we demonstrate identification of 1 000 protein groups for a standard HeLa cell line digest. The amount of loaded sample was varied in a range from 1 to 500 ng, and the method demonstrated 10-fold higher sensitivity. Combined with the recently introduced Diffacto approach for relative protein quantification, DirectMS1 outperforms most popular MS/MS-based label-free quantitation approaches because of significantly higher protein sequence coverage.
Keyphrases
- ms ms
- ultra high performance liquid chromatography
- tandem mass spectrometry
- liquid chromatography tandem mass spectrometry
- high performance liquid chromatography
- label free
- simultaneous determination
- mass spectrometry
- liquid chromatography
- high resolution
- amino acid
- small molecule
- binding protein
- drug delivery
- healthcare
- gas chromatography
- cancer therapy
- cell proliferation
- affordable care act
- signaling pathway
- cell death
- single cell
- wound healing