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Four Stage Liquid Chromatographic Selection of Methionyl Peptides for Peptide-Centric Proteome Analysis: The Proteome of Human Multipotent Adult Progenitor Cells
journal contribution
posted on 2006-06-02, 00:00 authored by Kris Gevaert, Jef Pinxteren, Hans Demol, Koen Hugelier, An Staes, Jozef Van Damme, Lennart Martens, Joël VandekerckhoveSerial application of strong cation-exchange and diagonal reversed-phase chromatography selecting
methionyl peptides by stepwise shifting them from their reduced to their sulfoxide and sulfone forms
generates a four-stage fractionation system, allowing high coverage analysis of complex proteome
digests by LC−MALDI−MS/MS. Application to the proteome of a human multipotent adult progenitor
cell line (MAPC) identified 2151 proteins with high confidence as on average four MS/MS-spectra were
linked to each protein. Our dataset contains several novel, potential marker proteins that may be
evaluated as affinity-anchors for isolating different adult stem cells in further studies. Furthermore, at
least 2 tyrosine kinases that were previously linked to the self-renewal potential of stem cells were
identified, validating the stemness of the analyzed cells. We also present data hinting at possible
involvement of the ubiquitin/proteasome machinery in steering proliferation and/or differentiation of
MAPC. Finally, following comparison of the MAPC proteome with proteomes of four human
differentiated cell lines reveals differential usage of chromosomal information: compared to differentiated cells, MAPC do not appear to hold any preference for expressing genes located on specific
chromosomes.
Keywords: diagonal chromatography • gel-free proteomics • COFRADIC • multidimensional chromatography
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multipotent adult progenitor cell linemethionyl peptideschromatographyMAPC proteomecell linesMethionyl Peptides2 tyrosine kinasesMSproteome digestscoverage analysisStage Liquid Chromatographic Selectionmarker proteinssulfone formsCOFRADIC2151 proteinsLCHuman Multipotent Adult Progenitor CellsSerial applicationsteering proliferation
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