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Titolo:
Oxidized forms of peroxiredoxins and DJ-1 on two-dimensional gels increased in response to sublethal levels of paraquat
Autore:
Mitsumoto, A; Nakagawa, Y; Takeuchi, A; Okawa, K; Iwamatsu, A; Takanezawa, Y;
Indirizzi:
Kitasato Univ, Sch Pharmaceut Sci, Tokyo 108, Japan Kitasato Univ Tokyo Japan 108 Univ, Sch Pharmaceut Sci, Tokyo 108, Japan Kirin Brewery Co Ltd, Cent Labs Key Technol, Yokohama, Kanagawa, Japan Kirin Brewery Co Ltd Yokohama Kanagawa Japan , Yokohama, Kanagawa, Japan
Titolo Testata:
FREE RADICAL RESEARCH
fascicolo: 3, volume: 35, anno: 2001,
pagine: 301 - 310
SICI:
1071-5762(2001)35:3<301:OFOPAD>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
OXIDATIVE STRESS; THIOREDOXIN PEROXIDASE; PLASMA ANTIOXIDANTS; PROTEIN OXIDATION; DNA-DAMAGE; MECHANISM; TOXICITY; GROWTH; CELLS; ELECTROPHORESIS;
Keywords:
hydroperoxide; peroxiredoxin; DJ-1; paraquat; 2D PAGE; endothelial cells;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Nakagawa, Y Kitasato Univ, Sch Pharmaceut Sci, 5-9-1 Shirokane, Tokyo 108,Japan Kitasato Univ 5-9-1 Shirokane Tokyo Japan 108 okyo 108, Japan
Citazione:
A. Mitsumoto et al., "Oxidized forms of peroxiredoxins and DJ-1 on two-dimensional gels increased in response to sublethal levels of paraquat", FREE RAD RE, 35(3), 2001, pp. 301-310

Abstract

We previously found hydroperoxide-responsive proteins (HPRPs), which are comprised of peroxiredoxin I (Prx I), Prx II, Prx III, Prx VI, HSP27, G3PDH and two unidentified proteins (HPRP-2' and HPRP-5'), in human umbilical vein endothelial cells. It was demonstrated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) that most HPRPs are converted into variants with lower pI upon exposure to hydroperoxides. In this study, we examined the HPRP response on 2D gels upon exposure of human endothelial cells (ECV304) to paraquat (PQ(2+)), which generates reactive oxygen species (ROS) within cells. PQ(2+) exerted cytotoxic effects in a dose- (10 muM-10 mM) and time- (24-168h) dependent manner. Two-dimensional PAGE analysis revealed that HPRP-2', and oxidized forms of Prx I, Prx II and Prx III were clearly increased upon exposure of cells to sublethal levels of PQ(2+). Microsequence analysis revealed that both HPRP-2 and -2' were identical with human DJ-1. Moreover immunoblot analysis confirmed the increase of oxidized forms of Prx II, Prx III and DJ-1 in response to sublethal levels of PQ(2+). PQ(2+) treatment failed to increase fluorescence intensity derived from DCF, which is believed to be an indicator for intracellular levels of hydroperoxide. Although pentachlorophenol (PCP), an uncoupler of the mitochondrial respiratorychain, clearly elevated the fluorescence, PCP had no effect on HPRP response. These observations indicated that DCF-derived fluorescence is not correlated with HPRP response. We consider that the response of Prxs and DJ-1 on2D gels could reflect endogenous production of ROS in PQ(2+)-treated cells, and might be a sensitive indicator of oxidative stress status.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/01/21 alle ore 03:34:47