Thioredoxin as a Molecular Target of Cyclopentenone Prostaglandins

Prostaglandin (PG) D2, a major cyclooxygenase product in a variety of tissues and cells, readily undergoes dehydration to yield the bioactive cyclopentenone-type PGs of the J2 series, such as 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2). We have shown previously that 15d-PGJ2 is a potent electrophile that causes intracellular oxidative stress and redox alteration in human neuroblastoma SH-SY5Y cells. In the present study, based on the observation that the electrophilic center of 15d-PGJ2 was involved in the pro-oxidant effect, we investigated the role of thioredoxin 1 (Trx), an endogenous redox regulator, against 15d-PGJ2-induced oxidative cell injury. It was observed that the 15d-PGJ2-induced oxidative stress was significantly suppressed by the Trx overexpression. In addition, the treatment of SH-SY5Y cells with biotinylated 15d-PGJ2 resulted in the formation of a 15d-PGJ2-Trx adduct, indicating that 15d-PGJ2 directly modified the endogenous Trx in the cells. To further examine the mechanism of the 15d-PGJ2 modification of Trx, human recombinant Trx treated with 15d-PGJ2 was analyzed by mass spectrometry. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the 15d-PGJ2-treated human recombinant Trx demonstrated the addition of one molecule of 15d-PGJ2 per protein molecule. Moreover, the electrospray ionization-liquid chromatography/mass spectrometry/mass spectrometry analysis identified two cysteine residues, Cys-35 and Cys-69, as the targets of 15d-PGJ2. These residues may represent the direct sensors of the electrophilic PGs that induce the intracellular redox alteration and neuronal cell death. Prostaglandin (PG) D2, a major cyclooxygenase product in a variety of tissues and cells, readily undergoes dehydration to yield the bioactive cyclopentenone-type PGs of the J2 series, such as 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2). We have shown previously that 15d-PGJ2 is a potent electrophile that causes intracellular oxidative stress and redox alteration in human neuroblastoma SH-SY5Y cells. In the present study, based on the observation that the electrophilic center of 15d-PGJ2 was involved in the pro-oxidant effect, we investigated the role of thioredoxin 1 (Trx), an endogenous redox regulator, against 15d-PGJ2-induced oxidative cell injury. It was observed that the 15d-PGJ2-induced oxidative stress was significantly suppressed by the Trx overexpression. In addition, the treatment of SH-SY5Y cells with biotinylated 15d-PGJ2 resulted in the formation of a 15d-PGJ2-Trx adduct, indicating that 15d-PGJ2 directly modified the endogenous Trx in the cells. To further examine the mechanism of the 15d-PGJ2 modification of Trx, human recombinant Trx treated with 15d-PGJ2 was analyzed by mass spectrometry. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the 15d-PGJ2-treated human recombinant Trx demonstrated the addition of one molecule of 15d-PGJ2 per protein molecule. Moreover, the electrospray ionization-liquid chromatography/mass spectrometry/mass spectrometry analysis identified two cysteine residues, Cys-35 and Cys-69, as the targets of 15d-PGJ2. These residues may represent the direct sensors of the electrophilic PGs that induce the intracellular redox alteration and neuronal cell death. The prostaglandins (PGs) 1The abbreviations used are: PG(s), prostaglandin(s); 15d-PGJ2, 15-deoxy-Δ12,14-PGJ2; Trx, thioredoxin; ELISA, enzyme-linked immunosorbent assay; ROS, reactive oxygen species; DCFH-DA, 2′,7′-dichlorodihydrofluorescein diacetate; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; HRP, horseradish peroxidase; PBS, phosphate-buffered saline; DTNB, 5,5′-dithiobis(2-nitrobenzoic acid); hrTrx, human recombinant Trx; ECL, enhanced chemiluminescence; HPLC, high performance liquid chromatography; TBS, Tris-buffered saline; MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight; MS, mass spectrometry; ESI-LC, electrospray ionization-liquid chromatography. are a family of structurally related molecules that are produced by cells in response to a variety of extrinsic stimuli and regulate cellular growth, differentiation, and homeostasis (1Smith W.L. Biochem. J. 1989; 259: 315-324Crossref PubMed Scopus (773) Google Scholar, 2Smith W.L. Am. J. Physiol. 1992; 263: F181-F191PubMed Google Scholar). PGs are derived from fatty acids, primarily arachidonate, which are released from membrane phospholipids by the action of phospholipases. Arachidonate is first converted to an unstable endoperoxide intermediate by cyclooxygenase and subsequently converted into one of several related products, including PGD2, PGE2, PGF2α, prostacyclin, and thromboxane A2, through the action of specific PG synthetases. Among them, PGD2 is a major cyclooxygenase product in a variety of tissues and cells and has marked effects on a number of biological processes, including platelet aggregation, relaxation of vascular and nonvascular smooth muscles, and nerve cell functions (3Giles H. Leff P. Prostaglandins. 1988; 35: 277-300Crossref PubMed Scopus (160) Google Scholar). It has been shown that PGD2 readily undergoes dehydration in vivo and in vitro to yield biologically active PGs of the J2 series, such as PGJ2, Δ12-PGJ2, and 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2) (4Fitzpatrick F.A. Wynalda M.A. J. Biol. Chem. 1983; 258: 11713-11718Abstract Full Text PDF PubMed Google Scholar, 5Kikawa Y. Narumiya S. Fukushima M. Wakatsuka H. Hayaishi O. Proc. Natl. Acad. U. S. A. 1984; 81: 1317-1321Crossref PubMed Scopus (166) Google Scholar, 6Hirata Y. Hayashi H. Ito S. Kikawa Y. Ishibashi M. Sudo M. Miyazaki H. Fukushima M. Narumiya S. Hayashi O. J. Biol. Chem. 1988; 263: 16619-16625Abstract Full Text PDF PubMed Google Scholar, 7Shibata T. Kondo M. Osawa T. Shibata N. Kobayashi M. Uchida K. J. Biol. Chem. 2002; 277: 10459-10466Abstract Full Text Full Text PDF PubMed Scopus (358) Google Scholar). Members of the J2 series of the PGs, unlike other classes of eicosanoids, characterized by the presence of an electrophilic α,β-unsaturated carbonyl group in the cyclopentenone ring, have their own unique spectrum of biological effects, including inhibition of macrophage-derived cytokine production (8Ricote M. Li A.C. Willson T.M. Kelly C.J. Glass C.K. Nature. 1998; 391: 79-82Crossref PubMed Scopus (3258) Google Scholar, 9Jiang C. Ting A.T. Seed B. Nature. 1998; 391: 82-86Crossref PubMed Scopus (538) Google Scholar) and IκB kinase (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar, 11Straus D. Pascual G. Li M. Welch J.S. Ricote M. Hsiang C.-H. Sengchanthalangsy L.L. Ghosh G. Glass C.K. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 4844-4849Crossref PubMed Scopus (947) Google Scholar), induction of synoviocyte and endothelial cell apoptosis (12Bishop-Bailey D. Hla T. J. Biol. Chem. 1998; 274: 17042-17048Abstract Full Text Full Text PDF Scopus (407) Google Scholar), induction of glutathione S-transferase gene expression (13Kawamoto Y. Nakamura Y. Naito Y. Torii Y. Kumagai T. Osawa T. Ohigashi H. Satoh K. Imagawa M. Uchida K. J. Biol. Chem. 2000; 275: 11291-11299Abstract Full Text Full Text PDF PubMed Scopus (95) Google Scholar) and intracellular oxidative stress (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar), and potentiation of apoptosis in neuronal cells (15Kondo M. Shibata T. Kumagai T. Osawa T. Shibata N. Kobayashi M. Sasaki S. Iwata M. Noguchi N. Uchida K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 7367-7372Crossref PubMed Scopus (163) Google Scholar). Moreover, recent studies have shown that 15d-PGJ2 directly inhibits the NF-κB-dependent gene expression through covalent modification of critical cysteine residues in IκB kinase (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar) and the DNA-binding domains of NF-κB subunits (11Straus D. Pascual G. Li M. Welch J.S. Ricote M. Hsiang C.-H. Sengchanthalangsy L.L. Ghosh G. Glass C.K. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 4844-4849Crossref PubMed Scopus (947) Google Scholar, 16Cernuda-Morollon E. Pineda-Molina E. Canada F.J. Perez-Sala D. J. Biol. Chem. 2001; 276: 35530-35536Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar). Thioredoxin 1 (Trx) is a small and ubiquitously expressed protein originally identified in Escherichia coli and is evolutionarily conserved from prokaryotes to higher eukaryotes (17Laurent T.C. Moore E.C. Reichard P. J. Biol. Chem. 1964; 239: 3436-3444Abstract Full Text PDF PubMed Google Scholar, 18Holmgren A. Annu. Rev. Biochem. 1985; 54: 237-271Crossref PubMed Google Scholar, 19Holmgren A. J. Biol. Chem. 1989; 264: 13963-13966Abstract Full Text PDF PubMed Google Scholar). Human thioredoxin was cloned as an adult T cell leukemia-derived factor or interleukin-1-like factor (20Tagaya Y. Y. A. Kondo N. H. J. N. K. T. H. J. 1989; PubMed Scopus Google Scholar, A. P. P. P. J. D. J. Biol. Chem. 1988; 263: Full Text PDF PubMed Google Scholar). Trx is an has other cellular including of and and as a redox active factor A. Annu. Rev. Biochem. 1985; 54: 237-271Crossref PubMed Google Scholar, A. J. Biochem. 2000; PubMed Scopus Google Scholar). In addition, Trx is to in the redox of and in against oxidative stress J. T. 1992; Full Text PDF PubMed Scopus Google Scholar, H. Nakamura K. J. Annu. Rev. PubMed Scopus Google Scholar). The of Trx in active the two cysteine residues and In addition to the conserved cysteine residues in the active cysteine residues Cys-69, and are present in the of the human The the action of Trx is that to a conserved in the of the the of as a with the protein to a the Cys-35 the the protein and which is by Trx A. Full Text Full Text PDF PubMed Scopus Google Scholar). In study, based on an of on the induction of intracellular production of reactive oxygen we identified cyclopentenone PGs, such as 15d-PGJ2, as the of intracellular oxidative stress in SH-SY5Y human neuroblastoma cells (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar). the intracellular with the oxidative we observed the cellular redox alteration by of such as glutathione and glutathione a in the membrane the production of products, such as and and the of In addition, the significantly the These that the redox alteration was related to the pro-oxidant of the cyclopentenone In the present study, to the the redox and 15d-PGJ2-induced oxidative stress and to the cellular mechanism against the endogenous we investigated the role of Trx in 15d-PGJ2-induced oxidative cell injury. and from and enhanced from The against and from and The protein was the protein from and from The modified was from Human recombinant Trx was produced by a previously Y. H. H. Nakamura H. Iwata S. A. S. N. T. J. PubMed Scopus Google Scholar) and by was from cells in and The cells in with and was by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium as previously (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar). of was used to the Google Scholar, G. G. J. Biol. PubMed Scopus Google Scholar). with in and treated with an on the cells with PBS, from the and 1 in The was a of group of 15d-PGJ2 was modified by with by a modification of a previously E. Pineda-Molina E. Canada F.J. Perez-Sala D. J. Biol. Chem. 2001; 276: 35530-35536Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar). 15d-PGJ2 was through a with a of The modified PG was and in further of in and The by the cells to The cells in and the The cells 1 in the presence of with and with of the cellular a laser with a and and was to Nature. PubMed Scopus Google Scholar). was a with and with was by the addition of or and The by with Trx in SH-SY5Y cells with or with the In 1 cells with of of in 1 of of 1 of was and cells by on of the cells by in of Trx in SH-SY5Y cells with biotinylated 15d-PGJ2 1 The cells with PBS, and in 1 of protein with of with The with by 1 The by the in and analyzed by by with In addition, the cell with of The was treated with of protein and 1 The was with and with the and the biotinylated to and with and the of the in was used and with of the biotinylated 15d-PGJ2 was to the by with was with of 1 The was to the 1 of and was and several in the The was by the addition of and the was on a of Trx with of the in was by with by the of Biochem. PubMed Scopus Google Scholar), as modified by B. 1983; PubMed Scopus Google Scholar). was treated with 15d-PGJ2, and against PBS, the protein was with and and with The was as an in The of was a with Matrix-assisted and with a of and and on targets and The with a of and and on targets and The an matrix-assisted laser desorption ionization time-of-flight mass with a laser in the and the was to series of on an mass with an electrospray The electrospray a and a of and with modified in of an of analyzed by a HPLC, a that of a with a a These with a of and The was and the was The and of to has been that the α,β-unsaturated carbonyl group in the cyclopentenone of 15d-PGJ2 is a the induction of intracellular oxidative stress and (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar). To human neuroblastoma SH-SY5Y cells to 15d-PGJ2 and and induction of production and shown in the intracellular production in SH-SY5Y cells was by 15d-PGJ2 in a The of in the cells to 15d-PGJ2 was higher that of the In to potent pro-oxidant of 15d-PGJ2, effects on the In a to the induction of the 15d-PGJ2 resulted in a in the to of the of the in the cells to the of the in the cyclopentenone of 15d-PGJ2 the pro-oxidant and effects of 15d-PGJ2, indicating that biological to the electrophilic center of 15d-PGJ2. effects of cyclopentenone-type PGs have been in several cell used or Google Scholar, C. M. T. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar, N. Biol. Google Scholar), we such in human neuroblastoma SH-SY5Y cells of Trx on that cyclopentenone PGs are to addition with that the action of the cyclopentenone PGs are related to the direct with glutathione other a specific of glutathione induce intracellular production and cell indicating that the effects of the cyclopentenone PGs may from glutathione (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar). and the that the electrophilic center of 15d-PGJ2 is involved in the that other cellular redox molecules in against 15d-PGJ2-induced oxidative cell injury. on the that Trx, a molecule in the of cellular redox critical in against oxidative stress and G. D. A. Biol. 2000; PubMed Scopus Google Scholar), we investigated the role of redox on the 15d-PGJ2-induced To we the of SH-SY5Y cells by with Trx The Trx expression was into the SH-SY5Y cells with the and with to a SH-SY5Y cells with a shown in the and demonstrated a readily expression of the Trx protein by analysis with the the cells and the shown the and to the of expressed Trx in against 15d-PGJ2-induced cell injury. we the intracellular production by shown in the production in the SH-SY5Y was significantly by 15d-PGJ2. The of in the cells to 15d-PGJ2 was higher that of the cells, the intracellular production in the cells to 15d-PGJ2 was of the we the of Trx on the 15d-PGJ2-induced The SH-SY5Y cells by to by to 15d-PGJ2. shown in 15d-PGJ2 or resulted in a in the to of the on cells of the cells to 15d-PGJ2-induced cell the cells. We the of Trx on the of and of which have been to involved in the 15d-PGJ2-induced cell (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar, M. Shibata T. Kumagai T. Osawa T. Shibata N. Kobayashi M. Sasaki S. Iwata M. Noguchi N. Uchida K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 7367-7372Crossref PubMed Scopus (163) Google Scholar), and that Trx have significantly the of These that Trx may in in the against the 15d-PGJ2 of to Trx in SH-SY5Y Trx reactive that Trx may directly with 15d-PGJ2. from adduct, the are with high The of IκB kinase (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar), Trx K. P. E. F.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), and M. Y. T. Y. T. K. K. and M. we the direct 15d-PGJ2 and endogenous Trx in SH-SY5Y cells. To 15d-PGJ2 with Trx or other we a biotinylated 15d-PGJ2 which the α,β-unsaturated and the electrophilic of 15d-PGJ2. shown in of the biotinylated 15d-PGJ2 into the cells was observed in SH-SY5Y cells by the of the biotinylated 15d-PGJ2. We to the 15d-PGJ2-Trx in the cells to the biotinylated 15d-PGJ2. To the SH-SY5Y cells treated with biotinylated 15d-PGJ2 1 and the cell was with with to the through biotinylated 15d-PGJ2 with and Trx was by analysis with the cell to with an and the presence of biotinylated was by analysis with that 15d-PGJ2 to an with endogenous Trx, in SH-SY5Y cells. of to Human of 15d-PGJ2 to Trx in vitro of with biotinylated 15d-PGJ2. shown in the biotinylated 15d-PGJ2 was to in a In addition, of the biotinylated 15d-PGJ2 to was shown by analysis Moreover, the 15d-PGJ2-treated with shown in the of to 15d-PGJ2 resulted in the of cysteine residues, and two cysteine residues per 1 These that 15d-PGJ2 the the of further the mechanism modification of Trx by 15d-PGJ2, we to the modification of the protein by the formation of the 15d-PGJ2 by mass shown in the analysis of the a of was with 15d-PGJ2 in subunits as was the to the addition of one molecule of 15d-PGJ2 per resulted in the of to the addition of one to two molecules of 15d-PGJ2 of the in the 15d-PGJ2 modification the and 15d-PGJ2-treated with and analyzed by mass by analysis of the from the of the of the protein and to the mass of the which an mass of to the addition of a molecule of 15d-PGJ2, and by analysis Moreover, two in the analysis of from the 15d-PGJ2-treated and identified by and from modification with 15d-PGJ2 in a modification with 15d-PGJ2 To the the and further analyzed by chromatography. The spectrum of the from the is shown in In the the product and and and The and product and the observed to that the 15d-PGJ2 modification is in the on Cys-35 The spectrum of the from the is shown in In the the product and and and and product and The product and and product and observed to These that the 15d-PGJ2 modification is with on the the other the modification of the other cysteine residues, and we identified the two cysteine residues, Cys-35 and Cys-69, of the 15d-PGJ2 modification in spectrum of the from the with the PGs are present in in to M. Google the is significantly in several including and Biol. PubMed Google Scholar), and PG in the have been of S. J. PubMed Scopus Google Scholar). In addition, cyclopentenone PG has been in the of D. J. PubMed Scopus Google Scholar). the of PG is to involved in the to The that PGD2, the of cyclopentenone PGs, is one of the produced PGs in several PGD2 readily converted to J2 PGs in the presence of in vitro Y. Narumiya S. Fukushima M. Wakatsuka H. Hayaishi O. Proc. Natl. Acad. U. S. A. 1984; 81: 1317-1321Crossref PubMed Scopus (166) Google and the cyclopentenone PGs, such as 15d-PGJ2, shown to from PGD2 in vivo Y. Hayashi H. Ito S. Kikawa Y. Ishibashi M. Sudo M. Miyazaki H. Fukushima M. Narumiya S. Hayashi O. J. Biol. Chem. 1988; 263: 16619-16625Abstract Full Text PDF PubMed Google Scholar), that the of the PGD2 may and related The of PGs in is by and which PG the of the in J. J. PubMed Scopus Google Scholar, U. M. G. A. Biol. Full Text PDF PubMed Scopus Google Scholar, J. J. M. Willson P. PubMed Google Scholar, PubMed Google Scholar, H. E. PubMed Google Scholar, E. G. M. PubMed Scopus Google Scholar). In addition, are on high of have a of A.T. J. 1989; PubMed Scopus Google Scholar, G. Chen 1992; PubMed Google Scholar, H. J. PubMed Scopus Google Scholar, C. J. PubMed Scopus Google Scholar). We have shown (15Kondo M. Shibata T. Kumagai T. Osawa T. Shibata N. Kobayashi M. Sasaki S. Iwata M. Noguchi N. Uchida K. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 7367-7372Crossref PubMed Scopus (163) Google Scholar) that 15d-PGJ2 is in the of in the of the is that through role in by the enhanced intracellular production of cyclopentenone PGs, is involved in In (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar), as of an to the endogenous of intracellular oxidative stress and to the mechanism the oxidative cell we the fatty their to induce intracellular production in SH-SY5Y cell in vitro and that the J2 series of the PGs the potent and the that the intracellular production was by the alteration of the cellular redox and the production of such as and which the intracellular that intracellular oxidative stress a in the of cellular by the In the present study, to further the mechanism the cell we the of an of 15d-PGJ2, on the induction of production and cell and that the of the in the cyclopentenone of 15d-PGJ2 the pro-oxidant and effects of 15d-PGJ2 the pro-oxidant action of 15d-PGJ2 to that the of electrophilic α,β-unsaturated The reactive center of the cyclopentenone PGs has been to of their biological (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar, T. 1998; PubMed Scopus Google Scholar). by of the addition with such as the of glutathione and cysteine residues in cellular that an role in the of the redox (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar, T. 1998; PubMed Scopus Google Scholar, M. 1992; Full Text PDF PubMed Scopus Google Scholar). in that cellular redox molecules in the of the biological functions of cyclopentenone In the present study, we investigated the role of Trx on the 15d-PGJ2-induced oxidative cell Trx, a molecule in the of the cellular redox has been shown to critical in against oxidative stress and G. Rev. 2001; PubMed Scopus Google Scholar). the of Trx against oxidative cell G. D. A. Biol. 2000; PubMed Scopus Google Scholar, A. G. Google Scholar). T. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar) have shown and as Trx the oxidative apoptosis of SH-SY5Y cells. with Trx on SH-SY5Y cells against 15d-PGJ2-induced cell It was that the of oxidative cell by Trx through a direct electrophile To a biotinylated 15d-PGJ2, we the of 15d-PGJ2 into cellular and cellular Trx that biotinylated 15d-PGJ2 Trx to cells. These the that Trx an role in the against the pro-oxidant effects of the electrophilic the in have that PGD2 and effects through with intracellular Narumiya S. K. Fukushima M. M. J. Google Scholar) have shown that is into cells and to the is with PGs, including PGD2, PGJ2, and Δ12-PGJ2, have been shown to with high to the fatty protein and intracellular protein involved in the intracellular and of fatty and their S. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). 15d-PGJ2 has been shown to directly NF-κB by IκB kinase through covalent of critical cysteine residues in IκB kinase or by with cysteine residues in the DNA-binding of the NF-κB (10Rossi A. Kapahi P. Natoli G. Takahashi T. Chen Y. Karin M. Santoro M.G. Nature. 2000; 403: 103-108Crossref PubMed Scopus (1202) Google Scholar, A. S. P. Biol. 2000; PubMed Scopus Google Scholar). Moreover, the NF-κB was identified to a covalent modification by 15d-PGJ2 to the inhibition of E. Pineda-Molina E. Canada F.J. Perez-Sala D. J. Biol. Chem. 2001; 276: 35530-35536Abstract Full Text Full Text PDF PubMed Scopus (284) Google Scholar). K. P. E. F.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) have the covalent modification and inhibition of Trx by cyclopentenone Perez-Sala D. A. N. Canada F.J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar) have shown that 15d-PGJ2 by direct of 15d-PGJ2 to the cysteine of and cyclopentenone PGs induce redox alteration in to the inhibition of the The covalent of 15d-PGJ2 to Trx was in vitro of with biotinylated 15d-PGJ2. The that the biotinylated 15d-PGJ2 was significantly into and that the of to the biotinylated 15d-PGJ2 resulted in the of cysteine residues that 15d-PGJ2 is a reactive of in the protein Trx In study, we have used a of and to the mechanism modification of Trx by 15d-PGJ2. The analysis of 15d-PGJ2-treated that the of Trx with 15d-PGJ2 resulted in the formation of two the major of which is a The 15d-PGJ2-Trx was further with and the and the adduct, by Among two cysteine residues and and other cysteine residues Cys-69, and in human Trx, the analysis the of modification to Cys-35 and These that 15d-PGJ2 may into may cysteine in specific The of Cys-35 to was the active of Trx is to have the on a with Cys-35 H. A. PubMed Scopus Google Scholar), and has a a Cys-35 H. C. C. PubMed Scopus Google Scholar). E. Chem. PubMed Scopus Google Scholar) that with the mechanism of Cys-35 by 15d-PGJ2 the modification of one of the two active may directly with the of redox Cys-35 is critical to the protein by the formation of a with the other the cysteine residues have been to covalent J. J. S. Biol. 2002; PubMed Scopus Google Scholar) that Trx is on Cys-69, which is an and a in the J.S. Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. A. Biol. 1998; PubMed Scopus Google Scholar). It has been that on is involved in an mechanism of Trx that from the by the of to and Cys-35 J. J. S. Biol. 2002; PubMed Scopus Google Scholar). is and the redox of Trx, the 15d-PGJ2 modification of may in an in the formation of may with (14Kondo M. Oya-Ito T. Kumagai T. Osawa T. Uchida K. J. Biol. Chem. 2001; 276: 12076-12083Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar) that 15d-PGJ2 and other cyclopentenone-type PGs induce intracellular oxidative stress in SH-SY5Y human neuroblastoma cells. In identified the Trx as a the covalent modification by 15d-PGJ2, a the redox alteration by cyclopentenone In addition, we identified the two cysteine residues, Cys-35 and Cys-69, of the 15d-PGJ2 modification The Trx modification by 15d-PGJ2 may one of the by which 15d-PGJ2 intracellular oxidative stress and neuronal cell death. We Kumagai

Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.