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Bhattacharjee A, et al.  (2009) In vivo protein tyrosine nitration in S. cerevisiae: Identification of tyrosine-nitrated proteins in mitochondria. Biochem Biophys Res Commun 388(3):612-7

Abstract: Protein tyrosine nitration (PTN) is a selective post translational modification often associated with pathophysiological conditions. Although yeast cells lack of mammalian nitric oxide synthase (NOS) orthologues, still it has been shown that they are capable of producing nitric oxide (NO). Our studies showed that NO or reactive nitrogen species (RNS) produced in flavohemoglobin mutant (Deltayhb1) strain along with the wild type strain (Y190) of S. cerevisiae can be visualized using specific probe 4, 5-diaminofluorescein diacetate (DAF-2DA). Deltayhb1 strain of S. cerevisiae showed bright fluorescence under confocal microscope that proves NO or RNS accumulation is more in absence of flavohemoglobin. We further investigated PTN profile of both cytosol and mitochondria of Y190 and Deltayhb1 cells of S. cerevisiae using two dimensional (2D) gel electrophoresis followed by western blot analysis. Surprisingly, we observed many immunopositive spots both in cytosol and in mitochondria from Y190 and Deltayhb1 using monoclonal anti-3-nitrotyrosine antibody indicating a basal level of NO or nitrite or peroxynitrite is produced in yeast system. To identify proteins nitrated in vivo we analyzed mitochondrial proteins from Y190 strains of S. cerevisiae. Among the eight identified proteins, two target mitochondrial proteins are aconitase and isocitrate dehydrogenase that are involved directly in the citric acid cycle. This investigation is the first comprehensive study to identify mitochondrial proteins nitrated in vivo.

Status: Published Type: Journal Article PubMed ID: 19695224

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ACO1 COQ6 FMP40 HEM15 IDH2 IML2 MAM33 TOM71 YHB1
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