Comprehensive Proteomic Analysis of Human Erythropoiesis

Abstract

SUMMARY Mass spectrometry-based proteomics now enables the absolute quantification of thousands of proteins in individual cell types. We used this technology to analyze the dynamic proteome changes occurring during human erythropoiesis. We quantified the absolute expression of 6,130 proteins during erythroid differentiation from late burst-forming units-erythroid (BFU-Es) to orthochromatic erythroblasts. A modest correlation between mRNA and protein expression was observed. We identified several proteins with unexpected expression patterns in erythroid cells, highlighting a breakpoint in the erythroid differentiation process at the basophilic stage. We also quantified the distribution of proteins between reticulocytes and pyrenocytes after enucleation. These analyses identified proteins that are actively sorted either with the reticulocyte or the pyrenocyte. Our study provides the absolute quantification of protein expression during a complex cellular differentiation process in humans, and it establishes a framework for future studies of disordered erythropoiesis.

Dynamic Regulation of a Ribosome Rescue Pathway in Erythroid Cells and Platelets

“…Strikingly however, PELO was initially up-regulated ~5-fold in the 2 days following hemin induction in K562 cells (Figure 3B–D) and more modestly in published quantitative mass spectrometry data from erythropoietic differentiation of primary CD34+ cells (Fig. 3F) (Gautier et al, 2016). …”

“…Indeed, we found that the levels of the canonical ribosome recycling factor ABCE1 (Pisarev et al, 2010) are strongly reduced in platelets compared to precursor megakaryoblast cells (Meg01) and in reticulocytes when compared to undifferentiated K562 cells (Figure S2C). Moreover, declining ABCE1 levels have been observed in proteomic analyses of erythroid differentiation from primary human cells (Gautier et al, 2016; Prenni et al, 2012), although the consequences of this decline were not explored. Interestingly, ABCE1 is also depleted in PLPs (Figure 3A) and in K562 cells induced toward erythroid differentiation (Figure 3B–C), where RNA levels also decline (Figure S4).…”

“…F . Quantitative mass spectrometry data from Gautier et al , 2016 showing relative protein levels during erythropoietic differentiation of primary CD34+ cells.…”

Enhanced calreticulin expression in red cells of polycythemia vera patients harboring the JAK2V617F mutation

“…We explored the expression of erythroid membrane proteins by performing a proteomic analysis of red blood cell ghosts from three PV patients and two age-matched controls (CT) using iTRAQ labeling technology. 8 Red blood cell ghosts were prepared by hypotonic lysis of washed cells as described elsewhere, 9 and peptides were prepared from 100 µg of total membrane proteins by trypsin digestion. Peptides from PV samples were labeled with 113, 115 and 117 iTRAQ reagents and peptides from CT samples with 118 and 121.…”

The Proteome of the Red Blood Cell: An Auspicious Source of New Insights into Membrane-Centered Regulation of Homeostasis

“…Many cytoplasmic proteins are reduced in number by enucleation and vesiculation during maturation of reticulocytes [29,30]. In addition, the process of aging-associated vesiculation results in the specific removal of both cytosplasmic proteins such as modified hemoglobins and damaged membrane proteins such as band 3 [31].…”