Tuesday, December 12, 2017 Print page
 
Centre of Inflammation and Metabolism (CIM)
 

Publications

To read the abstract, please click on the title of the publication of interest. If you want to access the publication on PubMed, please click on the PubMed ID.
To find specific publications, please use the sort and search functions. Please enter one word only as search term.

Click here to see all publications

191Stable isotope labeling by amino acids in cell culture (SILAC) and quantitative comparison of the membrane proteomes of self-renewing and differentiating human embryonic stem cells.
Prokhorova TA; Rigbolt KT; Johansen PT; Henningsen J; Kratchmarova I; Kassem M; Blagoev B
Mol Cell Proteomics 2009; 8(5): 959-70
PubMed ID: 19151416

Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful quantitative proteomics platform for comprehensive characterization of complex biological systems. However, the potential of SILAC-based approaches has not been fully utilized in human embryonic stem cell (hESC) research mainly because of the complex nature of hESC culture conditions. Here we describe complete SILAC labeling of hESCs with fully preserved pluripotency, self-renewal capabilities, and overall proteome status that was quantitatively analyzed to a depth of 1556 proteins and 527 phosphorylation events. SILAC-labeled hESCs appear to be perfectly suitable for functional studies, and we exploited a SILAC-based proteomics strategy for discovery of hESC-specific surface markers. We determined and quantitatively compared the membrane proteomes of the self-renewing versus differentiating cells of two distinct human embryonic stem cell lines. Of the 811 identified membrane proteins, six displayed significantly higher expression levels in the undifferentiated state compared with differentiating cells. This group includes the established marker CD133/Prominin-1 as well as novel candidates for hESC surface markers: Glypican-4, Neuroligin-4, ErbB2, receptor-type tyrosine-protein phosphatase zeta (PTPRZ), and Glycoprotein M6B. Our study also revealed 17 potential markers of hESC differentiation as their corresponding protein expression levels displayed a dramatic increase in differentiated embryonic stem cell populations. Prokhorova, Tatyana A



 
© 2017 Centre of Inflammation and Metabolism
www.inflammation-metabolism.dk