proteomic analysis of erk12-mediated human sickle red blood cell membrane protein phosphorylation人类蛋白质组学分析erk12-mediated镰状血红细胞膜蛋白质磷酸化.pdfVIP

Soderblom et al. Clinical Proteomics 2013, 10:1 /content/10/1/1 CLINICAL PROTEOMICS RESEARCH Open Access Proteomic analysis of ERK1/2-mediated human sickle red blood cell membrane protein phosphorylation 1 1 2 2 1 1 Erik J Soderblom , J Will Thompson , Evan A Schwartz , Edward Chiou , Laura G Dubois , M Arthur Moseley and Rahima Zennadi2,3* Abstract Background: In sickle cell disease (SCD), the mitogen-activated protein kinase (MAPK) ERK1/2 is constitutively active and can be inducible by agonist-stimulation only in sickle but not in normal human red blood cells (RBCs). ERK1/2 is involved in activation of ICAM-4-mediated sickle RBC adhesion to the endothelium. However, other effects of the ERK1/2 activation in sickle RBCs leading to the complex SCD pathophysiology, such as alteration of RBC hemorheology are unknown. Results: To further characterize global ERK1/2-induced changes in membrane protein phosphorylation within human RBCs, a label-free quantitative phosphoproteomic analysis was applied to sickle and normal RBC membrane ghosts pre-treated with U0126, a specific inhibitor of MEK1/2, the upstream kinase of ERK1/2, in the presence or absence of recombinant active ERK2. Across eight unique treatment groups, 375 phosphopeptides from 155 phosphoproteins were quantified with an average technical coefficient of variation in peak intensity of 19.8%. Sickle RBC treatment with U0126 decreased thirty-six phosphopeptides from twenty-one phosphoproteins involved in regulation of not only RBC shape, flexibility, cell morphol