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

2.50
Hdl Handle:
http://hdl.handle.net/10541/72742
Title:
Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells.
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
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.
Affiliation:
Stem Cell and Leukaemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Manchester M20 4QL, UK.
Citation:
Quantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells. 2006, 107 (12):4687-94 Blood
Journal:
Blood
Issue Date:
15-Jun-2006
URI:
http://hdl.handle.net/10541/72742
DOI:
10.1182/blood-2005-12-4995
PubMed ID:
16507774
Type:
Article
Language:
en
ISSN:
0006-4971
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorUnwin, Richard D-
dc.contributor.authorSmith, Duncan L-
dc.contributor.authorBlinco, David-
dc.contributor.authorWilson, Claire L-
dc.contributor.authorMiller, Crispin J-
dc.contributor.authorEvans, Caroline A-
dc.contributor.authorJaworska, Ewa-
dc.contributor.authorBaldwin, Stephen A-
dc.contributor.authorBarnes, Kay-
dc.contributor.authorPierce, Andrew-
dc.contributor.authorSpooncer, Elaine-
dc.contributor.authorWhetton, Anthony D-
dc.date.accessioned2009-07-07T11:52:32Z-
dc.date.available2009-07-07T11:52:32Z-
dc.date.issued2006-06-15-
dc.identifier.citationQuantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells. 2006, 107 (12):4687-94 Blooden
dc.identifier.issn0006-4971-
dc.identifier.pmid16507774-
dc.identifier.doi10.1182/blood-2005-12-4995-
dc.identifier.urihttp://hdl.handle.net/10541/72742-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.subjectHaematopoietic Stem Cellsen
dc.subject.meshAnimals-
dc.subject.meshCell Hypoxia-
dc.subject.meshChromatography, Liquid-
dc.subject.meshHematopoietic Stem Cells-
dc.subject.meshMass Spectrometry-
dc.subject.meshMice-
dc.subject.meshOxidation-Reduction-
dc.subject.meshProtein Processing, Post-Translational-
dc.subject.meshProteome-
dc.subject.meshProteomics-
dc.titleQuantitative proteomics reveals posttranslational control as a regulatory factor in primary hematopoietic stem cells.en
dc.typeArticleen
dc.contributor.departmentStem Cell and Leukaemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Manchester M20 4QL, UK.en
dc.identifier.journalBlooden

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