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A proteomic analysis of CBA‐Pk‐1slc/Pk‐1slc mice with Red Blood Cell Type Pyruvate Kinase deficiency using 2DE together with MALDI–TOF‐MS analysis
Author(s) -
Kedar Prabhakar,
Shimizu Kazuyuki,
Ghosh Kanjaksha,
Fujji Hisaichi,
Colah Roshan,
Kanno Hitoshi
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1025.1
Subject(s) - pyruvate kinase , chemistry , heat shock protein , oxidative stress , proteome , peptide mass fingerprinting , erythropoietin , cytosol , downregulation and upregulation , proteomics , apoptosis , hsp70 , oxidative phosphorylation , kinase , glycolysis , microbiology and biotechnology , biochemistry , biology , gene , enzyme , endocrinology
SLC3 is a Friend erythroleukemic cells line from the Pk‐1slc mouse erythrocyte pyruvate kinase(PK)deficiency. SLC3 undergo spontaneous apoptosis leads to ineffective erythropoiesis. It was showed that glycolytic inhibition by PKLR gene mutation augmented oxidative stress, leading to the activation of HIF‐1 as well as downstream proapoptotic gene expression. We studied proteomic response of SLC3 was quantitatively analyzed two‐dimensional gel electrophoresis (2DE) followed by identification with matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI‐TOF‐MS). We detected significant differences between SLC3 and a control Friend cell. 72% of the total proteins studied were significantly down regulated in SLC3 and only 28% of proteins were up regulated. Several membrane proteins are significantly down‐regulated in SLC3 may cause reduced stability and integrity of PK deficient RBC. Oxidative stress proteins in SLC3 mice were up‐regulated and antioxidant proteins were down regulated. These proteins related to generation of ROS. For the refolding of mutant protein, molecular chaperon is expectedly unregulated in SLC3. Upregulation of two chaperons are presumably an adapted reaction in SLC3. Heat‐shock protein (HSP70) and mortalin, A novel mediator of erythropoietin signaling. We speculated that down‐regulation of mortalin might be attributable to disturbance of EPO signaling, resulting in premature death of cells. The proteomic approach makes an important contribution to characterizing the proteome of the SLC3.