Limits...
Phosphoproteomic analysis of apoptotic hematopoietic stem cells from hemoglobin E/β-thalassemia.

Ponnikorn S, Panichakul T, Sresanga K, Wongborisuth C, Roytrakul S, Hongeng S, Tungpradabkul S - J Transl Med (2011)

Bottom Line: Ineffective erythropoiesis has been studied in human hematopoietic stem cells, however the distinct apoptotic mechanism was unclear.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.

ABSTRACT

Background: Hemoglobin E/β-thalassemia is particularly common in Southeast Asia and has variable symptoms ranging from mild to severe anemia. Previous investigations demonstrated the remarkable symptoms of β-thalassemia 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/β-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/β-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/β-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/β-thalassemic stem cells is an effective method to further investigate the cell death mechanism of ineffective erythropoiesis in β-thalassemia. Our report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/β-thalassemia.

Show MeSH

Related in: MedlinePlus

Schematic diagram of candidate phosphoproteins implicated in apoptotic pathways. The color pictures represent proteins that were identified in this study, the grey pictures are proteins that have been previously identified.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3142509&req=5

Figure 4: Schematic diagram of candidate phosphoproteins implicated in apoptotic pathways. The color pictures represent proteins that were identified in this study, the grey pictures are proteins that have been previously identified.

Mentions: Interestingly, Several proteins that were over-expressed in patients were identified as apoptotic proteins. Of these, five proteins were characterized as common in the apoptotic pathway including cytochrome C, apoptosis inducing factor 1 (AIFM1), caspase 6, tumor necrosis factor ligand 6 (TNFL 6) and tumor necrosis factor receptor super family 12A (TNFR 12A or TWEAK). These proteins were involved in both mitochondrial dependent apoptosis (intrinsic mechanism) and death receptor mediated apoptosis (extrinsic mechanism). Previous evidence suggests that the pathology and disease progression of β-thalassemia relates to the accumulation of reactive oxygen species (ROS), which can generate DNA adducts in the nucleus and induce the DNA damage pathway. In this study, six proteins encoded by four genes were identified and found to be responsible for DNA damage and responsive mechanisms such as the p53 pathway. Interestingly, we found the isoforms sigma, zeta/delta and gamma of the 14-3-3 protein were over-expressed in patients more than 2 fold as compared with normal donors. These 14-3-3 proteins are involved in cellular pathways including apoptosis and phosphatidyl inositol 3 kinase activity by PANTHER pathway analysis. A schematic diagram of the apoptotic pathway including the identified phosphoproteins in HbE/β-thalassemic stem cells was proposed (Figure 4).


Phosphoproteomic analysis of apoptotic hematopoietic stem cells from hemoglobin E/β-thalassemia.

Ponnikorn S, Panichakul T, Sresanga K, Wongborisuth C, Roytrakul S, Hongeng S, Tungpradabkul S - J Transl Med (2011)

Schematic diagram of candidate phosphoproteins implicated in apoptotic pathways. The color pictures represent proteins that were identified in this study, the grey pictures are proteins that have been previously identified.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3142509&req=5

Figure 4: Schematic diagram of candidate phosphoproteins implicated in apoptotic pathways. The color pictures represent proteins that were identified in this study, the grey pictures are proteins that have been previously identified.
Mentions: Interestingly, Several proteins that were over-expressed in patients were identified as apoptotic proteins. Of these, five proteins were characterized as common in the apoptotic pathway including cytochrome C, apoptosis inducing factor 1 (AIFM1), caspase 6, tumor necrosis factor ligand 6 (TNFL 6) and tumor necrosis factor receptor super family 12A (TNFR 12A or TWEAK). These proteins were involved in both mitochondrial dependent apoptosis (intrinsic mechanism) and death receptor mediated apoptosis (extrinsic mechanism). Previous evidence suggests that the pathology and disease progression of β-thalassemia relates to the accumulation of reactive oxygen species (ROS), which can generate DNA adducts in the nucleus and induce the DNA damage pathway. In this study, six proteins encoded by four genes were identified and found to be responsible for DNA damage and responsive mechanisms such as the p53 pathway. Interestingly, we found the isoforms sigma, zeta/delta and gamma of the 14-3-3 protein were over-expressed in patients more than 2 fold as compared with normal donors. These 14-3-3 proteins are involved in cellular pathways including apoptosis and phosphatidyl inositol 3 kinase activity by PANTHER pathway analysis. A schematic diagram of the apoptotic pathway including the identified phosphoproteins in HbE/β-thalassemic stem cells was proposed (Figure 4).

Bottom Line: Ineffective erythropoiesis has been studied in human hematopoietic stem cells, however the distinct apoptotic mechanism was unclear.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.

ABSTRACT

Background: Hemoglobin E/β-thalassemia is particularly common in Southeast Asia and has variable symptoms ranging from mild to severe anemia. Previous investigations demonstrated the remarkable symptoms of β-thalassemia 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/β-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/β-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/β-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/β-thalassemic stem cells is an effective method to further investigate the cell death mechanism of ineffective erythropoiesis in β-thalassemia. Our report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/β-thalassemia.

Show MeSH
Related in: MedlinePlus