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Alternative NADH dehydrogenase (NDH2): intermembrane-space-facing counterpart of mitochondrial complex I in the procyclic Trypanosoma brucei
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SYSNO ASEP 0392793 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Alternative NADH dehydrogenase (NDH2): intermembrane-space-facing counterpart of mitochondrial complex I in the procyclic Trypanosoma brucei Tvůrce(i) Verner, Zdeněk (BC-A)
Škodová, Ingrid (BC-A) ORCID
Poláková, S. (CZ)
Ďurišová-Benkovičková, V. (SK)
Horváth, A. (SK)
Lukeš, Julius (BC-A) RID, ORCIDZdroj.dok. Parasitology. - : Cambridge University Press - ISSN 0031-1820
Roč. 140, č. 3 (2013), s. 328-337Poč.str. 10 s. Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova Trypanosoma ; mitochondrion ; dehydrogenase ; respiration ; NDH2 Vědní obor RIV EB - Genetika a molekulární biologie CEP LC07032 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy GA204/09/1667 GA ČR - Grantová agentura ČR Institucionální podpora BC-A - RVO:60077344 UT WOS 000315341200006 DOI 10.1017/S003118201200162X Anotace The respiratory chain of the procyclic stage of Trypanosoma brucei contains the standard complexes I through IV, as well as several alternative enzymes contributing to electron flow. In this work, we studied the function of an alternative NADH: ubiquinone oxidoreductase (NDH2). Depletion of target mRNA was achieved using RNA interference (RNAi). In the non-induced and RNAi-induced cell growth, membrane potential change, alteration in production of reactive oxygen species, overall respiration, enzymatic activities of complexes I, III and/or IV and distribution of NADH: ubiquinone oxidoreductase activities in glycerol gradient fractions were measured. Finally, respiration using different substrates was tested on digitonin-permeabilized cells. The induced RNAi cell line exhibited slower growth, decreased mitochondrial membrane potential and lower sensitivity of respiration to inhibitors. Mitochondrial glycerol-3-phosphate dehydrogenase was the only enzymatic activity that has significantly changed in the interfered cells. This elevation as well as a decrease of respiration usingNADHwas confirmed on digitonin-permeabilized cells. The data presented here together with previously published findings on complex I led us to propose thatNDH2 is the majorNADH: ubiquinone oxidoreductase responsible for cytosolic and not for mitochondrial NAD+ regeneration in the mitochondrion of procyclic T. brucei. Pracoviště Biologické centrum (od r. 2006) Kontakt Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Rok sběru 2014
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