A two-dimensional electrophoretic map of human mitochondrial proteins from immortalized lymphoblastoid cell lines: A prerequisite to study mitochondrial disorders in patients

Xie, Jing; Techritz, Sandra; Haebel, Sophie; Horn, Anke; Neitzel, Heidemarie; Klose, Joachim; Schuelke, Markus

doi:10.1002/pmic.200401191

Cited by 21 publications

(20 citation statements)

References 58 publications

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“…To date, the proteome analysis of mitochondria includes studies in human heart (Hprot_01?06) [35][36][37], mouse mitochondria (various tissues, only proteins with clear human orthologs (Hprot_02) [38], human placenta (Hprot_03) [39], human lymphoblastoid cell lines (Hprot_04) [40], neuroblastoma cell lines (Hprot_05) [41], and rat liver (Rprot_01) [42]. Four additional highthroughput analyses performed in the mouse have been added.…”

Section: Human and Mousementioning

confidence: 98%

MitoP2: An Integrative Tool for the Analysis of the Mitochondrial Proteome

et al. 2008

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Mitochondria are crucial for normal cell metabolism and maintenance. Mitochondrial dysfunction has been implicated in a spectrum of human diseases, ranging from rare monogenic to common multifactorial disorders. Important for the understanding of organelle function is the assignment of its constituents, and although over 1,500 proteins are predicted to be involved in mammalian mitochondrial function, so far only about 900 are assigned to mitochondria with reasonable certainty. Continuing efforts are being taken to obtain a complete inventory of the mitochondrial proteome by single protein studies and high-throughput approaches. To be of best value for the scientific community this data needs to be structured, explored, and customized. For this purpose, the MitoP2 database ( http://www.mitop2.de ) was established and is maintained in order to incorporate such data. The central database contains manually evaluated yeast, mouse, and human reference proteins, which show convincing evidence of a mitochondrial location. In addition, entries from genome-wide approaches that suggest protein localization are integrated and serve to compile a combined score for each candidate, which provides a best estimate of mitochondrial localization. Furthermore, it integrates information on the orthology between species, including Saccharomyces cerevisiae, mouse, human, Arabidopsis thaliana, and Neurospora crassa, thus mutually enhancing evidence across species. In contrast to other known databases, MitoP2 takes into account the reliability by which the protein is estimated as being mitochondrially located, as described herein. Multiple search functions, as well as information on disease causing genes and available mouse models, makes MitoP2 a valuable tool for the genetic investigation of human mitochondrial pathology.

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Section: Human and Mousementioning

confidence: 98%

MitoP2: An Integrative Tool for the Analysis of the Mitochondrial Proteome

et al. 2008

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“…Competent Escherichia coli (JM 109 cells) were then transformed with the plasmids containing the PCR product inserts. White colonies (8)(9)(10)(11)(12), including recombinant plasmids, were selected and cultured. After plasmid extraction, length heteroplasmic mutations were determined by DNA sequencing.…”

Section: Determination Of Mtdna Copy Numbermentioning

confidence: 99%

“…Purification of mitochondria, sample preparation, and run for two-dimensional gel electrophoresis (2-DE) were performed using the previous published protocols [9]. Coomassie blue and silver-stained gels were digitized using a densitometer (ImageScanner), and analyzed with PDQuest 7.3.1 computer software (Bio-Rad).…”

Section: Proteomics Of Mitochondrial-rich Cytoplasmic Fractionmentioning

confidence: 99%

Mitochondrial DNA copy number and hnRNP A2/B1 protein: Biomarkers for direct exposure of benzene

Eom

Kim

et al. 2011

Enviro Toxic and Chemistry

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The present study was performed to identify biomarkers for exposure of benzene in blood cells and hematopoietic tissues. Peripheral mononuclear cells, hematopoietic stem cells, and leukemia cell lines were cultured in RPMI 1640 media with the addition of 0, 1, and 10 mM of benzene. Hydrogen peroxide was measured using an enzyme immunoassay. Mitochondrial mass, membrane potential, and mitochondrial DNA (mtDNA) copy number were measured using MitoTracker Green/Red probes, and real-time polymerase chain reaction. In addition, two-dimensional gel electrophoresis and mass spectrometry matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) technology were performed to identify protein markers. The mitochondrial contents and membrane potentials were dramatically increased after three weeks of direct benzene exposure. The hydrogen peroxide level increased significantly after two weeks of treatment with benzene (4.4 ± 1.9 µM/mg protein) compared to the non-benzene treatment group (1.2 ± 1.0; p = 0.001). The mtDNA copy number gradually increased after exposure to benzene. Numerous protein markers showed significant aberrant expression after exposure to benzene. Among them, the heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1 was markedly decreased after exposure to benzene. Thus, increased mitochondrial mass, mtDNA copy number, and the hnRNP A2/B1 protein were biomarkers for benzene-related toxicity and hematotoxicity.

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“…Even though the human mitochondrial genome has been well characterized [34], a description of the mitochondrial sub-proteome [35][36][37][38][39] is lacking in many human tissues, including skeletal muscle, in spite of the relevance of this information to understanding the molecular basis of mitochondrial myopathies (for a review see [40][41][42]). …”

Section: References 6 Addendummentioning

confidence: 99%

Coordinated and reversible reduction of enzymes involved in terminal oxidative metabolism in skeletal muscle mitochondria from a riboflavin‐responsive, multiple acyl‐CoA dehydrogenase deficiency patient

et al. 2006

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In this case report we studied alterations in mitochondrial proteins in a patient suffering from recurrent profound muscle weakness, associated with ethylmalonic-adipic aciduria, who had benefited from high dose of riboflavin treatment. Morphological and biochemical alterations included muscle lipid accumulation, low muscle carnitine content, reduction in fatty acid beta-oxidation and reduced activity of complexes I and II of the respiratory chain. Riboflavin therapy partially or totally reversed these symptoms and increased the level of muscle flavin adenine dinucleotide, suggesting that aberrant flavin cofactor metabolism accounted for the disease. Proteomic investigation of muscle mitochondria revealed decrease or absence of several flavoenzymes, enzymes related to flavin cofactor-dependent mitochondrial pathways and mitochondrial or mitochondria-associated calcium-binding proteins. All these deficiencies were completely rescued after riboflavin treatment. This study indicates for the first time a profound involvement of riboflavin/flavin cofactors in modulating the level of a number of functionally coordinated polypeptides involved in fatty acyl-CoA and amino acid metabolism, extending the number of enzymatic pathways altered in riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency.

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A two-dimensional electrophoretic map of human mitochondrial proteins from immortalized lymphoblastoid cell lines: A prerequisite to study mitochondrial disorders in patients

Cited by 21 publications

References 58 publications

MitoP2: An Integrative Tool for the Analysis of the Mitochondrial Proteome

MitoP2: An Integrative Tool for the Analysis of the Mitochondrial Proteome

Mitochondrial DNA copy number and hnRNP A2/B1 protein: Biomarkers for direct exposure of benzene

Coordinated and reversible reduction of enzymes involved in terminal oxidative metabolism in skeletal muscle mitochondria from a riboflavin‐responsive, multiple acyl‐CoA dehydrogenase deficiency patient

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