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Title: Phosphoproteomic analysis of apoptotic hematopoietic stem cells from hemoglobin E/b-thalassemia
Authors: Saranyoo Ponnikorn
Tasanee Panichakul
Kitima Sresanga
Chokdee Wongborisuth
Sittiruk Roytrakul
Suradej Hongeng
Sumalee Tungpradabkul
Mahidol University. Faculty of Science. Department of Biochemistry
Keywords: Open Access article;Phosphoproteome;Hemoglobin E/β-thalassemia;HSCs/CD34+;apoptosis
Issue Date: 2011
Citation: Journal of Translational Medicine. Vol. 9, (2011), 96
Abstract: Background: Hemoglobin E/b-thalassemia is particularly common in Southeast Asia and has variable symptoms ranging from mild to severe anemia. Previous investigations demonstrated the remarkable symptoms of bthalassemia in terms of the acceleration of apoptotic cell death. Ineffective erythropoiesis has been studied in human hematopoietic stem cells, however the distinct apoptotic mechanism was unclear. Methods: The phosphoproteome of bone marrow HSCs/CD34+ cells from HbE/b-thalassemic patients was analyzed using IMAC phosphoprotein isolation followed by LC-MS/MS detection. Decyder MS software was used to quantitate differentially expressed proteins in 3 patients and 2 normal donors. The differentially expressed proteins from HSCs/CD34+ cells were compared with HbE/b-thalassemia and normal HSCs. Results: A significant change in abundance of 229 phosphoproteins was demonstrated. Importantly, the analysis of the candidate proteins revealed a high abundance of proteins that are commonly found in apoptotic cells including cytochrome C, caspase 6 and apoptosis inducing factors. Moreover, in the HSCs patients a significant increase was observed in a specific type of phosphoserine/threonine binding protein, which is known to act as an important signal mediator for the regulation of cell survival and apoptosis in HbE/b-thalassemia. Conclusions: Our study used a novel method to investigate proteins that influence a particular pathway in a given disease or physiological condition. Ultimately, phosphoproteome profiling in HbE/b-thalassemic stem cells is an effective method to further investigate the cell death mechanism of ineffective erythropoiesis in b-thalassemia. Our report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/b-thalassemia.
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