Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells.
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
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
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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.
Description
Date
2006-06-15
Publisher
Collections
Keywords
Haematopoietic Stem Cells
Type
Article
Citation
Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells. 2006, 107 (12):4687-94 Blood