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Comparative proteomics of primitive hematopoietic cell populations reveals differences in expression of proteins regulating motility
- Source :
- Blood; May 2004, Vol. 103 Issue: 10 p3751-3759, 9p
- Publication Year :
- 2004
-
Abstract
- Lineage-marker depleted (Lin–) murine bone marrow cells expressing stem cell antigen 1 (Sca-1) were sorted on the basis of stem cell factor receptor (c-kit) expression to obtain Lin–Sca+Kit+ or Lin–Sca+Kit– cells. Lin–Sca+Kit– cells have a markedly greater chemotactic response to stromal derived factor-1 (SDF-1). Using a novel fluorescent stain, we show that both populations generate similar levels of a key messenger, phosphatidylinositol 3,4,5 trisphosphate (PIP3), in response to SDF-1. Differences in motile behavior may therefore lie downstream of phosphatidylinositol 3-kinase (PI3-kinase) activation at the level of cytoskeleton regulation. The 2 cell populations were compared using 2-dimensional difference gel electrophoresis (2D-DIGE), with a maleimide CyDye fluorescent protein labeling technique that has enhanced sensitivity for low abundance samples. Comparative proteomic analysis of Cy3- and Cy5-labeled protein samples allows relative quantification of protein spots present in both cell populations; of these, 73% were common. Key protein differences were adseverin and gelsolin, actin micro-filament splicing proteins, regulated by Rac, downstream of PI3-kinase activation. Adseverin was shown to be acetylated, a novel modification for this protein. Differences in major regulators of cell shape and motility between the 2 populations can explain the differential response to SDF-1.
Details
- Language :
- English
- ISSN :
- 00064971 and 15280020
- Volume :
- 103
- Issue :
- 10
- Database :
- Supplemental Index
- Journal :
- Blood
- Publication Type :
- Periodical
- Accession number :
- ejs10689217
- Full Text :
- https://doi.org/10.1182/blood-2003-09-3294