Document Detail


Plasma membrane proteomics of human embryonic stem cells and human embryonal carcinoma cells.
MedLine Citation:
PMID:  18489135     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human embryonic stem cells (hESCs) are of immense interest in regenerative medicine as they can self-renew indefinitely and can give rise to any adult cell type. Human embryonal carcinoma cells (hECCs) are the malignant counterparts of hESCs found in testis tumors. hESCs that have acquired chromosomal abnormalities in culture are essentially indistinguishable from hECC. Direct comparison of karyotypically normal hESCs with hECCs could lead to understanding differences between their mechanisms of growth control and contribute to implementing safe therapeutic use of stem cells without the development of germ cell cancer. While several comparisons of hECCs and hESCs have been reported, their cell surface proteomes are largely unknown, partly because plasma membrane proteomics is still a major challenge. Here, we present a strategy for the identification of plasma membrane proteins that has been optimized for application to the relatively small numbers of stem cells normally available, and that does not require tedious cell fractionation. The method led to the identification of 237 and 219 specific plasma membrane proteins in the hESC line HUES-7 and the hECC line NT2/D1, respectively. In addition to known stemness-associated cell surface markers like ALP, CD9, and CTNNB, a large number of receptors, transporters, signal transducers, and cell-cell adhesion proteins were identified. Our study revealed that several Hedgehog and Wnt pathway members are differentially expressed in hESCs and hECCs including NPC1, FZD2, FZD6, FZD7, LRP6, and SEMA4D, which play a pivotal role in stem cell self-renewal and cancer growth. Various proteins encoded on chromosome 12p, duplicated in testicular cancer, were uniquely identified in hECCs. These included GAPDH, LDHB, YARS2, CLSTN3, CSDA, LRP6, NDUFA9, and NOL1, which are known to be upregulated in testicular cancer. Distinct HLA molecules were revealed on the surface of hESCs and hECCs, despite their low abundance. Results were compared with genomic and proteomic data sets reported previously for mouse ESCs, hECCs, and germ cell tumors. Our data provides a surface signature for HUES-7 and NT2/D1 cells and distinguishes normal hESCs from hECCs, helping explain their 'benign' versus 'malignant' nature.
Authors:
Wilma Dormeyer; Dennis van Hoof; Stefan R Braam; Albert J R Heck; Christine L Mummery; Jeroen Krijgsveld
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-05-20
Journal Detail:
Title:  Journal of proteome research     Volume:  7     ISSN:  1535-3893     ISO Abbreviation:  J. Proteome Res.     Publication Date:  2008 Jul 
Date Detail:
Created Date:  2008-07-07     Completed Date:  2008-09-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128775     Medline TA:  J Proteome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2936-51     Citation Subset:  IM    
Affiliation:
Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Membrane / metabolism
Chromatography, Liquid
Embryonal Carcinoma Stem Cells / metabolism
Embryonic Stem Cells / metabolism*
Gene Expression Profiling
Humans
Membrane Proteins / genetics,  metabolism*
Mice
Organelles / metabolism
Proteomics
Species Specificity
Tandem Mass Spectrometry
Chemical
Reg. No./Substance:
0/Membrane Proteins

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