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Blood, 15 May 2004, Vol. 103, No. 10, pp. 3751-3759. Prepublished online as a Blood First Edition Paper on February 5, 2004; DOI 10.1182/blood-2003-09-3294. This Article Full Text (PDF) All Versions of this Article: 2003-09-3294v1 103/10/3751    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Evans, C. A Articles by Whetton, A. D Search for Related Content PubMed PubMed Citation Articles by Evans, C. A Articles by Whetton, A. D Related Collections Related Article in Blood Online Social Bookmarking                   What's this? Submitted October 3, 2003 Accepted January 27, 2004 Comparative proteomics of primitive hematopoietic cell populations reveals differences in expression of proteins regulating motility Caroline A Evans, Robert Tonge, David Blinco, Andrew Pierce, Joanne Shaw, Yuning Lu, Hajja G Hamzah, Alexander Gray, C P Downes, Simon J Gaskell, Elaine Spooncer, and Anthony D Whetton* Leukaemia Research Fund Proteomics Facility, UMIST, Manchester, United Kingdom Global Protein Science & Supply, Enabling Science & Technology (Biology), Astra Zeneca, Alderley Edge, Cheshire, United Kingdom LRF Cellular Development Unit, UMIST, Manchester, United Kingdom School of Life Sciences, University of Dundee, Dundee, United Kingdom Leukaemia Research Fund Proteomics Facility, UMIST, Manchester, United Kingdom; Michael Barber Centre for Mass Spectrometry, Dept of Chemistry, UMIST, Manchester, United Kingdom Leukaemia Research Fund Proteomics Facility, UMIST, Manchester, United Kingdom; LRF Cellular Development Unit, UMIST, Manchester, United Kingdom * Corresponding author; email: tony.whetton{at}umist.ac.uk. 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 behaviour may therefore lie downstream of phosphatidylinositol 3-kinase (PI 3-Kinase) activation at the level of cytoskeleton regulation. The two cell populations were compared using two-dimensional difference gel electrophoresis (2D-DIGE), with a maleimide CyDyeTM fluorescent protein labelling technique that has enhanced sensitivity for low abundance samples. Comparative proteomic analysis of Cy3 and Cy5 labelled protein samples allows relative quantification of protein spots present in the both cell populations and of these 73% were common. Key protein differences were adseverin and gelsolin, actin microfilament splicing proteins; regulated by Rac, downstream of PI 3-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 two populations can explain the differential response to SDF-1. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Proteomics knocks on hematology's door Beerelli Seshi Blood 2004 103: 3607. [Full Text] [PDF] This article has been cited by other articles: A. Pierce, R. D. Unwin, C. A. Evans, S. Griffiths, L. Carney, L. Zhang, E. Jaworska, C.-F. Lee, D. Blinco, M. J. Okoniewski, et al. Eight-channel iTRAQ Enables Comparison of the Activity of Six Leukemogenic Tyrosine Kinases Mol. Cell. Proteomics, May 1, 2008; 7(5): 853 - 863. [Abstract] [Full Text] [PDF] A. J. K. Williamson, D. L. Smith, D. Blinco, R. D. Unwin, S. Pearson, C. Wilson, C. Miller, L. Lancashire, G. Lacaud, V. Kouskoff, et al. Quantitative Proteomics Analysis Demonstrates Post-transcriptional Regulation of Embryonic Stem Cell Differentiation to Hematopoiesis Mol. Cell. Proteomics, March 1, 2008; 7(3): 459 - 472. [Abstract] [Full Text] [PDF] R. D. Unwin, D. L. Smith, D. Blinco, C. L. Wilson, C. J. Miller, C. A. Evans, E. Jaworska, S. A. Baldwin, K. Barnes, A. Pierce, et al. Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells Blood, June 15, 2006; 107(12): 4687 - 4694. [Abstract] [Full Text] [PDF] Y. Hu, J. P. Malone, A. M. Fagan, R. R. Townsend, and D. M. Holtzman Comparative Proteomic Analysis of Intra- and Interindividual Variation in Human Cerebrospinal Fluid Mol. Cell. Proteomics, December 1, 2005; 4(12): 2000 - 2009. [Abstract] [Full Text] [PDF] A. Kolkman, E. H. C. Dirksen, M. Slijper, and A. J. R. Heck Double Standards in Quantitative Proteomics: Direct Comparative Assessment of Difference in Gel Electrophoresis and Metabolic Stable Isotope Labeling Mol. Cell. Proteomics, March 1, 2005; 4(3): 255 - 266. [Abstract] [Full Text] [PDF] R. D. Unwin, D. W. Sternberg, Y. Lu, A. Pierce, D. G. Gilliland, and A. D. Whetton Global Effects of BCR/ABL and TEL/PDGFR{beta} Expression on the Proteome and Phosphoproteome: IDENTIFICATION OF THE RHO PATHWAY AS A TARGET OF BCR/ABL J. Biol. Chem., February 25, 2005; 280(8): 6316 - 6326. [Abstract] [Full Text] [PDF]

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