A two-dimensional electrophoretic map of human mitochondrial proteins from immortalized lymphoblastoid cell lines: a prerequisite to study mitochondrial disorders in patients |
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Authors: | Xie Jing Techritz Sandra Haebel Sophie Horn Anke Neitzel Heidemarie Klose Joachim Schuelke Markus |
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Institution: | Department of Neuropediatrics, Charité, University Medical Center, Berlin, Germany. |
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Abstract: | Mitochondrial diseases may be caused by numerous mutations that alter proteins of the respiratory chain and of other metabolic pathways in the mitochondrium. For clinicians this disease group poses a considerable diagnostic challenge due to ambiguous genotype-phenotype relationships. Until now, only 30% of the mitochondriopathies can be diagnosed at the molecular level. We therefore need a new diagnostic tool that offers a wide view on the mitochondrial proteins. Here, we present a method to generate a high-resolution, large-gel two-dimensional gel electrophoretic (2-DE) map of a purified fraction of mitochondrial proteins from Epstein-Barr virus-immortalized lymphoblastoid cell line (LCL). LCLs can be easily obtained from patients and control subjects in a routine clinical setting. They often express the biochemical phenotype and can be cultured to high cell numbers, sufficient to gain enough purified material for 2-DE. In total we identified 166 mitochondrial proteins. Thirteen proteins were earlier not known to be of mitochondrial origin. Thirty-nine proteins were associated with human diseases ranging from respiratory chain enzyme deficiencies to disorders of beta-oxidation and amino acid metabolism. This 2-DE map is intended to be the first step to diagnose mitochondrial diseases at the proteomic level. |
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Keywords: | Epstein‐Barr virus‐immortalized lymphoblastoid cells Leigh syndrome Matrix assisted laser desorption/ionization‐time of flight mass spectrometry/mass spectrometry Mitochondrial proteins Protein expression map Two‐dimensional gel electrophoresis |
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