In early stages of Alzheimer's disease (AD), amyloid beta (Aβ) accumulates in the mitochondrial matrix and interacts with mitochondrial proteins, such as cyclophilin D (cypD) and 17β-hydroxysteroid dehydrogenase 10 (17β-HSD10). Multiple processes associated with AD such as increased production or oligomerization of Aβ affect these interactions and disbalance the equilibrium between the biomolecules, which contributes to mitochondrial dysfunction. Here, we investigate the effect of the ionic environment on the interactions of Aβ (Aβ1-40, Aβ1-42) with cypD and 17β-HSD10 using a surface plasmon resonance (SPR) biosensor. We show that changes in concentrations of K+ and Mg2+ significantly affect the interactions and may increase the binding efficiency between the biomolecules by up to 35% and 65% for the interactions with Aβ1-40 and Aβ1-42, respectively, in comparison with the physiological state. We also demonstrate that while the binding of Aβ1-40 to cypD and 17β-HSD10 takes place preferentially around the physiological concentrations of ions, decreased concentrations of K+ and increased concentrations of Mg2+ promote the interaction of both mitochondrial proteins with Aβ1-42. These results suggest that the ionic environment represents an important factor that should be considered in the investigation of biomolecular interactions taking place in the mitochondrial matrix under physiological as well as AD-associated conditions.

• MeSH
• 17-hydroxysteroidní dehydrogenasy chemie   genetika   MeSH
• Alzheimerova nemoc diagnóza   genetika   patologie   MeSH
• amyloidní beta-protein chemie   MeSH
• biosenzitivní techniky metody   MeSH
• ionty chemie   MeSH
• lidé MeSH
• mitochondriální proteiny chemie   MeSH
• mitochondrie chemie   MeSH
• peptidové fragmenty chemie   genetika   MeSH
• peptidylprolylisomerasa F chemie   genetika   MeSH
• povrchová plasmonová rezonance metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A-kinase interacting protein 1 (AKIP1) has been shown to interact with a broad range of proteins involved in various cellular processes, including apoptosis, tumorigenesis, and oxidative stress suggesting it might have multiple cellular functions. In this study, we used an epitope-tagged AKIP1 and by combination of immunochemical approaches, microscopic methods and reporter assays we studied its properties. Here, we show that various levels of AKIP1 overexpression in HEK-293 cells affected not only its subcellular localization but also resulted in aggregation. While highly expressed AKIP1 accumulated in electron-dense aggregates both in the nucleus and cytosol, low expression of AKIP1 resulted in its localization within the nucleus as a free, non-aggregated protein. Even though AKIP1 was shown to interact with p65 subunit of NF-kappaB and activate this transcription factor, we did not observe any effect on NF-kappaB activation regardless of various AKIP1 expression level.

• MeSH
• adaptorové proteiny signální transdukční biosyntéza   genetika   MeSH
• buněčné jádro chemie   metabolismus   MeSH
• cytosol chemie   metabolismus   MeSH
• HEK293 buňky MeSH
• jaderné proteiny biosyntéza   genetika   MeSH
• lidé MeSH
• mitochondrie chemie   metabolismus   MeSH
• NF-kappa B analýza   metabolismus   MeSH
• regulace genové exprese MeSH
• subcelulární frakce chemie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Emerging evidence has linked mitochondrial dysfunction to the pathogenesis of many renal disorders, including acute kidney injury, sepsis and even chronic kidney disease. Proteomics is a powerful tool in elucidating the role of mitochondria in renal pathologies. Since the pig is increasingly recognized as a major mammalian model for translational research, the lack of physiological proteome data of large mammals prompted us to examine renal mitochondrial proteome in porcine kidney cortex and medulla METHODS: Kidneys were obtained from six healthy pigs. Mitochondria from cortex and medulla were isolated using differential centrifugation and proteome maps of cortical and medullar mitochondria were constructed using two-dimensional gel electrophoresis (2DE). Protein spots with significant difference between mitochondrial fraction of renal cortex and medulla were identified by mass spectrometry. RESULTS: Proteomic analysis identified 81 protein spots. Of these spots, 41 mitochondrial proteins were statistically different between renal cortex and medulla (p < 0.05). Protein spots containing enzymes of beta oxidation, amino acid metabolism, and gluconeogenesis were predominant in kidney cortex mitochondria. Spots containing tricarboxylic acid cycle enzymes and electron transport system proteins, proteins maintaining metabolite transport and mitochondrial translation were more abundant in medullar mitochondria. CONCLUSION: This study provides the first proteomic profile of porcine kidney cortex and medullar mitochondrial proteome. Different protein expression pattern reflects divergent functional metabolic role of mitochondria in various kidney compartments. Our study could serve as a useful reference for further porcine experiments investigating renal mitochondrial physiology under various pathological states.

• MeSH
• 2D gelová elektroforéza MeSH
• dřeň ledvin chemie   MeSH
• kůra ledviny chemie   MeSH
• mitochondriální proteiny analýza   MeSH
• mitochondrie chemie   MeSH
• modely u zvířat MeSH
• proteomika metody   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• Sus scrofa MeSH
• tandemová hmotnostní spektrometrie MeSH
• translační biomedicínský výzkum metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Iron-sulphur clusters (ISCs) are protein co-factors essential for a wide range of cellular functions. The core iron-sulphur cluster assembly machinery resides in the mitochondrion, yet due to export of an essential precursor from the organelle, it is also needed for cytosolic and nuclear iron-sulphur cluster assembly. In mitochondria all [4Fe-4S] iron-sulphur clusters are synthesised and transferred to specific apoproteins by so-called iron-sulphur cluster targeting factors. One of these factors is the universally present mitochondrial Nfu1, which in humans is required for the proper assembly of a subset of mitochondrial [4Fe-4S] proteins. Although most eukaryotes harbour a single Nfu1, the genomes of Trypanosoma brucei and related flagellates encode three Nfu genes. All three Nfu proteins localise to the mitochondrion in the procyclic form of T. brucei, and TbNfu2 and TbNfu3 are both individually essential for growth in bloodstream and procyclic forms, suggesting highly specific functions for each of these proteins in the trypanosome cell. Moreover, these two proteins are functional in the iron-sulphur cluster assembly in a heterologous system and rescue the growth defect of a yeast deletion mutant.

• MeSH
• chemická frakcionace MeSH
• down regulace MeSH
• elektroporace MeSH
• fylogeneze MeSH
• kultivované buňky MeSH
• mitochondriální proteiny fyziologie   MeSH
• mitochondrie chemie   fyziologie   MeSH
• plazmidy MeSH
• proteiny obsahující železo a síru genetika   imunologie   fyziologie   MeSH
• proteiny tepelného šoku HSP70 metabolismus   MeSH
• protilátky protozoální biosyntéza   MeSH
• protozoální proteiny genetika   imunologie   fyziologie   MeSH
• RNA interference MeSH
• Trypanosoma brucei brucei chemie   klasifikace   genetika   fyziologie   MeSH
• výpočetní biologie MeSH
• western blotting MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND AND AIMS: Dysfunction of kidney mitochondria plays a critical role in the pathogenesis of a number of renal diseases. Proteomics represents an untargeted attempt to reveal the remodeling of mitochondrial proteins during disease. Combination of separation methods and mass spectrometry allows identification and quantitative analysis of mitochondrial proteins including protein complexes. The aim of this review is to summarize the methods and applications of proteomics to renal mitochondria. METHODS: Using keywords "mitochondria", "kidney", "proteomics", scientific databases (PubMed and Web of knowledge) were searched from 2000 to August 2015 for articles describing methods and applications of proteomics to analysis of mitochondrial proteins in kidney. Included were publications on mitochondrial proteins in kidneys of humans and animal model in health and disease. RESULTS AND CONCLUSION: Proteomics of renal mitochondria has been/is mostly used in diabetes, hypertension, acidosis, nephrotoxicity and renal cancer. Integration of proteomics with other methods for examining protein activity is promising for insight into the role of renal mitochondria in pathological states. Several challenges were identified: selection of appropriate model organism, sensitivity of analytical methods and analysis of mitochondrial proteome in different renal zones/biopsies in the course of various kidney disorders.

• MeSH
• acidóza etiologie   MeSH
• akutní poškození ledvin etiologie   MeSH
• diabetické nefropatie etiologie   MeSH
• elektroforéza metody   MeSH
• ledviny chemie   MeSH
• lidé MeSH
• mitochondriální proteiny analýza   MeSH
• mitochondrie chemie   fyziologie   MeSH
• nádory ledvin etiologie   MeSH
• nemoci ledvin etiologie   MeSH
• proteomika metody   MeSH
• renální hypertenze etiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The human 5'(3')-deoxyribonucleotidases catalyze the dephosphorylation of deoxyribonucleoside monophosphates to the corresponding deoxyribonucleosides and thus help to maintain the balance between pools of nucleosides and nucleotides. Here, the structures of human cytosolic deoxyribonucleotidase (cdN) at atomic resolution (1.08 Å) and mitochondrial deoxyribonucleotidase (mdN) at near-atomic resolution (1.4 Å) are reported. The attainment of an atomic resolution structure allowed interatomic distances to be used to assess the probable protonation state of the phosphate anion and the side chains in the enzyme active site. A detailed comparison of the cdN and mdN active sites allowed the design of a cdN-specific inhibitor.

• MeSH
• cytosol chemie   enzymologie   MeSH
• deoxyribonukleotidy chemie   MeSH
• Escherichia coli genetika   metabolismus   MeSH
• eukaryotické buňky chemie   enzymologie   MeSH
• fosfáty chemie   MeSH
• inhibitory enzymů chemie   MeSH
• izoenzymy antagonisté a inhibitory   chemie   genetika   MeSH
• katalytická doména MeSH
• konformace proteinů MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• mitochondrie chemie   enzymologie   MeSH
• molekulární modely MeSH
• nukleotidasy antagonisté a inhibitory   chemie   genetika   MeSH
• organofosfonáty chemie   MeSH
• orgánová specificita MeSH
• racionální návrh léčiv MeSH
• rekombinantní proteiny chemie   genetika   MeSH
• simulace molekulového dockingu MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cardiolipin, an anionic phospholipid, in the inner mitochondrial membrane is involved in the regulation of mitochondrial bioenergetics and in oxidative phosphorylation. In addition, cardiolipin plays also some role in mitochondria-dependent steps of apoptosis and in mitochondrial membrane dynamics. Alterations in cardiolipin structure (remodelling), its content or the composition of acyl chains as the consequences of oxidative damage due to reactive oxygen species (ROS) are proposed to be responsible for the changes in the mitochondrial membrane fluidity, ion permeability and structure/function of the mitochondrial electron-transport chain components. The mitochondrial dysfunction caused by the above events has been associ¬ated with several physiopathological conditions in human tissues, including Barth syndrome, ischemia/reperfusion, different thyroid states, diabetes, aging or heart failure.

• MeSH
• apoptóza MeSH
• Barthův syndrom etiologie   MeSH
• biogeneze organel * MeSH
• buněčná membrána fyziologie   MeSH
• Ca2+-ATPasy MeSH
• fosfolipidy biosyntéza   fyziologie   MeSH
• homeostáza MeSH
• kardiolipiny * biosyntéza   fyziologie   metabolismus   MeSH
• kardiovaskulární nemoci etiologie   MeSH
• lidé MeSH
• mitochondriální ADP/ATP-translokasy MeSH
• mitochondriální protonové ATPasy MeSH
• mitochondrie * fyziologie   chemie   MeSH
• nemoci endokrinního systému etiologie   MeSH
• neurodegenerativní nemoci etiologie   MeSH
• oxidační stres MeSH
• peroxidace lipidů MeSH
• reaktivní formy kyslíku MeSH
• Check Tag
• lidé MeSH

Coenzyme Q (CoQ), a lipophilic substituted benzoquinone present in all cells. Besides its fundamental role of an electron carrier associated with energy production in the respiratory chain, CoQ has two other functions in mitochondria. It is an essential factor in activation of protein uncoupling and it controls permeability of transition pores. Moreover, it participates in extramitochondrial electron transport in plasma membranes and lysosomes, controls physicochemical properties of membranes, and is the only endogenous lipid antioxidant. Its pro-oxidant role consists in generating the major superoxide radical/H2O2 secondmessenger system. Biosynthesis of CoQ proceeds in every cell, small amounts of CoQ can be obtained from diet. CoQ is also available as a dietary supplement. It shows minimal toxicity, excellent tolerance, and no significant side effects. Its beneficial effects are largely attributed to its essential role in cellular bioenergetics and antioxidant properties. Supplementation of CoQ can improve conditions of a wide range of pathological states. Some forms of mitochondrial CoQ deficiency respond well to its oral administration. Recent meta-analysis of tests for hypertension is also promising. This review summarizes the current knowledge of the therapeutic efficacy of CoQ in various diseases.

• MeSH
• antioxidancia terapeutické užití   MeSH
• energetický metabolismus fyziologie   účinky léků   MeSH
• financování organizované MeSH
• klinické zkoušky jako téma využití   MeSH
• lékové interakce fyziologie   MeSH
• lidé MeSH
• medicína založená na důkazech trendy   MeSH
• mitochondrie fyziologie   chemie   metabolismus   MeSH
• nežádoucí účinky léčiv metabolismus   MeSH
• oxidace-redukce MeSH
• oxidancia izolace a purifikace   MeSH
• potravní doplňky využití   MeSH
• ubichinon dějiny   izolace a purifikace   terapeutické užití   MeSH
• Check Tag
• lidé MeSH

The KlPGS1 gene encoding phosphatidylglycerolphosphate synthase (PGPS) is essential for the viability and multiplication of Kluyveromyces lactis. Regulation of PGPS expression by factors affecting mitochondrial development (C source, growth phase) and general phospholipid biosynthesis was followed. PGS1 mRNA levels were not altered as cells progressed from the exponential to the stationary phase of growth in glucose. PGS1 mRNA abundance was nearly identical in cells growing in a medium with glucose or glycerol as the sole C source during the different growth phases. Regulation of PGS1 expression by exogenous myo-inositol and choline was not mediated at the transcriptional level, the PGPS activity dropped to 70 % after myo-inositol addition.

• MeSH
• aerobióza MeSH
• cholin metabolismus   MeSH
• fungální proteiny genetika   chemie   metabolismus   MeSH
• genetická transkripce MeSH
• inositol metabolismus   MeSH
• Kluyveromyces MeSH
• mitochondrie genetika   chemie   metabolismus   MeSH
• regulace genové exprese enzymů MeSH
• regulace genové exprese u hub MeSH
• terciární struktura proteinů MeSH
• transferasy pro jiné substituované fosfátové skupiny MeSH

• Klíčová slova
• Provinol,
• MeSH
• alfa-tokoferol chemie   MeSH
• fenoly farmakologie   MeSH
• finanční podpora výzkumu jako téma MeSH
• hypertenze farmakoterapie   MeSH
• krevní tlak účinky záření   MeSH
• mitochondrie chemie   MeSH
• mozek MeSH
• potkani inbrední SHR MeSH
• synthasa oxidu dusnatého chemie   MeSH
• ubichinon chemie   MeSH
• víno MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH