Four stage liquid chromatographic selection of methionyl peptides for peptide-centric proteome analysis: The proteome of human multipotent adult progenitor cells

被引：23

作者：

Gevaert, K ^{[1
]}

Pinxteren, J ^{[1
]}

Demol, H ^{[1
]}

Hugelier, K ^{[1
]}

Staes, A ^{[1
]}

Van Damme, J ^{[1
]}

Martens, L ^{[1
]}

Vandekerckhove, J ^{[1
]}

机构：

[1] Univ Ghent, Fac Med & Hlth Sci, Dept Biochem & Med Prot Res, B-9000 Ghent, Belgium

来源：

JOURNAL OF PROTEOME RESEARCH | 2006年 / 5卷 / 06期

关键词：

diagonal chromatography; gel-free proteomics; COFRADIC; multidimensional chromatography;

D O I：

10.1021/pr060026a

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Serial 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.

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收藏

页码：1415 / 1428

页数：14

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