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    Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells.

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    Authors
    Unwin, Richard D
    Smith, Duncan L
    Blinco, David
    Wilson, Claire L
    Miller, Crispin J
    Evans, Caroline A
    Jaworska, Ewa
    Baldwin, Stephen A
    Barnes, Kay
    Pierce, Andrew
    Spooncer, Elaine
    Whetton, Anthony D
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    Affiliation
    Stem Cell and Leukaemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Manchester M20 4QL, UK.
    Issue Date
    2006-06-15
    
    Metadata
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    Abstract
    The proteome is determined by rates of transcription, translation, and protein turnover. Definition of stem cell populations therefore requires a stem cell proteome signature. However, the limit to the number of primary cells available has restricted extensive proteomic analysis. We present a mass spectrometric method using an isobaric covalent modification of peptides for relative quantification (iTRAQ), which was employed to compare the proteomes of approximately 1 million long-term reconstituting hematopoietic stem cells (Lin(-)Sca(+)Kit(+); LSK(+)) and non-long-term reconstituting progenitor cells (Lin(-)Sca(+)Kit(-); LSK(-)), respectively. Extensive 2-dimensional liquid chromatography (LC) peptide separation prior to mass spectrometry (MS) enabled enhanced proteome coverage with relative quantification of 948 proteins. Of the 145 changes in the proteome, 54% were not seen in the transcriptome. Hypoxia-related changes in proteins controlling metabolism and oxidative protection were observed, indicating that LSK(+) cells are adapted for anaerobic environments. This approach can define proteomic changes in primary samples, thereby characterizing the molecular signature of stem cells and their progeny.
    Citation
    Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells. 2006, 107 (12):4687-94 Blood
    Journal
    Blood
    URI
    http://hdl.handle.net/10541/72742
    DOI
    10.1182/blood-2005-12-4995
    PubMed ID
    16507774
    Type
    Article
    Language
    en
    ISSN
    0006-4971
    ae974a485f413a2113503eed53cd6c53
    10.1182/blood-2005-12-4995
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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