Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells.
Authors
Unwin, Richard DSmith, 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
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
Show full item recordAbstract
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 BloodJournal
BloodDOI
10.1182/blood-2005-12-4995PubMed ID
16507774Type
ArticleLanguage
enISSN
0006-4971ae974a485f413a2113503eed53cd6c53
10.1182/blood-2005-12-4995
Scopus Count
Collections
Related articles
- Large-Scale and Deep Quantitative Proteome Profiling Using Isobaric Labeling Coupled with Two-Dimensional LC-MS/MS.
- Authors: Gritsenko MA, Xu Z, Liu T, Smith RD
- Issue date: 2016
- Application of targeted mass spectrometry in bottom-up proteomics for systems biology research.
- Authors: Manes NP, Nita-Lazar A
- Issue date: 2018 Oct 30
- Sample Preparation Approaches for iTRAQ Labeling and Quantitative Proteomic Analyses in Systems Biology.
- Authors: Spanos C, Moore JB
- Issue date: 2016
- Quantitative proteomic analysis using isobaric protein tags enables rapid comparison of changes in transcript and protein levels in transformed cells.
- Authors: Unwin RD, Pierce A, Watson RB, Sternberg DW, Whetton AD
- Issue date: 2005 Jul
- Hypoxia mediates low cell-cycle activity and increases the proportion of long-term-reconstituting hematopoietic stem cells during in vitro culture.
- Authors: Eliasson P, Rehn M, Hammar P, Larsson P, Sirenko O, Flippin LA, Cammenga J, Jönsson JI
- Issue date: 2010 Apr