Developmental fate determination and marker discovery in hematopoietic stem cell biology using proteomic fingerprinting.
In hematopoiesis, co-expression of Sca-1 and c-Kit defines cells (LS(+)K) with long term reconstituting potential. In contrast, poorly characterized LS(-)K cells fail to reconstitute lethally irradiated recipients. Relative quantification mass spectrometry and transcriptional profiling were used to characterize LS(+)K and LS(-)K cells. This approach yielded data on >1200 proteins. Only 32% of protein changes correlated to mRNA modulation demonstrating post-translational protein regulation in early hematopoietic development. LS(+)K cells had lower expression of protein synthesis proteins but did express proteins associated with mature cell function. Major increases in erythroid development proteins were observed in LS(-)K cells; based on this assessment of erythroid potential we showed them to be principally erythroid progenitors, demonstrating effective use of discovery proteomics for definition of primitive cells.
Spooncer
Elaine
E
Stem Cell and Leukaemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Kinnaird House, Kinnaird Road, Manchester M20 4QL, United Kingdom.
Brouard
Nathalie
N
Nilsson
Susie K
SK
Williams
Brenda
B
Liu
Mira C
MC
Unwin
Richard D
RD
Blinco
David
D
Jaworska
Ewa
E
Simmons
Paul J
PJ
Whetton
Anthony D
AD
D000818:Animals; D000067696:Ataxin-1; D000067528:Ataxins; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D019070:Cell Lineage; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D010641:Phenotype; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
C000593749:ATXN1 protein, human; D000067696:Ataxin-1; D000067528:Ataxins; C000593750:Atxn1 protein, mouse; D015415:Biomarkers; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D020543:Proteome; D012333:RNA, Messenger; D019009:Proto-Oncogene Proteins c-kit