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Research

Developmental Fate Determination and Marker Discovery in Hematopoietic Stem Cell Biology Using Proteomic Fingerprinting^*,S

Elaine Spooncer, Nathalie Brouard^,¶, Susie K. Nilsson^,||, Brenda Williams^,||, Mira C. Liu, Richard D. Unwin, David Blinco, Ewa Jaworska, Paul J. Simmons^,¶ and Anthony D. Whetton^,**

From the Stem Cell and Leukaemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Kinnaird House, Kinnaird Road, Manchester M20 4QL, United Kingdom and Stem Cell Laboratory, Peter MacCallum Cancer Centre, St. Andrews Place, East Melbourne, Victoria 3002, Australia

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.