Expression of Human Phospholipid Hydroperoxide Glutathione Peroxidase Gene for Protection of Host Cells from Lipid Hydroperoxide-Mediated Injury

Yagi, Kazuichi; Komura, Sadaaki; Kojima, Hiroharu; Sun, Qian; Nagata, Noriko; Ohishi, Nobuko; Nishikimi, Morimitsu

doi:10.1006/bbrc.1996.0260

Cited by 72 publications

(47 citation statements)

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“…For example, the overexpression of Gpx4 in RBL2H3 and 104C1 cells has been shown to reduce ROS-induced toxicity (33,34). Gpx4, especially mitochondrial Gpx4, also has been suggested to play a role in the protection against apoptosis.…”

Section: Discussionmentioning

confidence: 99%

Transgenic Mice Overexpressing Glutathione Peroxidase 4 Are Protected against Oxidative Stress-induced Apoptosis

Ran

Liang

et al. 2004

Journal of Biological Chemistry

229

192

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Glutathione peroxidase 4 (Gpx4) is uniquely involved in the detoxification of oxidative damage to membrane lipids. Our previous studies showed that Gpx4 is essential for mouse survival and that Gpx4 deficiency makes cells vulnerable to oxidative injury. In the present study, we generated two lines of transgenic mice overexpressing Gpx4 (Tg(GPX4) mice) using a genomic clone containing the human GPX4 gene. Both lines of Tg-(GPX4) mice, Tg5 and Tg6, had elevated levels of Gpx4 (mRNA and protein) in all tissues investigated, and overexpression of Gpx4 did not cause alterations in activities of glutathione peroxidase 1, catalase, Cu/Zn superoxide dismutase, and manganese superoxide dismutase. The human GPX4 transgene rescued the lethal phenotype of null mutation of the mouse Gpx4 gene, indicating that the transgene can replace the essential role of mouse Gpx4 in mouse development. Cell death induced by t-butylhydroperoxide and diquat was significantly less in murine embryonic fibroblasts from Tg(GPX4) mice compared with wild type mice. Liver damage and lipid peroxidation induced by diquat were reduced significantly in Tg(GPX4) mice. In addition, diquat-induced apoptosis was decreased in Tg(GPX4) mice, as evidenced by attenuated caspase-3 activation and reduced cytochrome c release from mitochondria. These data demonstrate that Gpx4 plays a role in vivo in the mechanism of apoptosis induced by oxidative stress that most likely occurs through oxidative damage to mitochondrial phospholipids such as cardiolipin.Reactive oxygen species (ROS), 1 such as superoxide and hydrogen peroxide, are constantly generated in aerobic organisms during normal respiration. In addition, environmental factors (such as ionizing radiation) and pathological compounds (such as ␤-amyloid in Alzheimer's disease) can generate ROS. Although ROS at physiological concentrations may be required for normal cell function, excessive production of ROS can be detrimental to cells, because ROS can cause oxidative damage to lipids, protein, and DNA. Polyunsaturated fatty acids, which are found predominantly in cellular membranes, are especially vulnerable to attack by ROS because of the high concentration of allylic hydrogens in their structure (1). The resulting lipid hydroperoxides can affect membrane fluidity and the function of membrane proteins. In addition, lipid hydroperoxides can undergo iron-mediated, one-electron reduction and oxygenation to form epoxyallylic peroxyl radicals, which trigger a chain reaction of free radical-mediated lipid peroxidation (2). The end-products of lipid peroxidation are reactive aldehydes such as 4-hydroxyl nonenal and malondialdehyde, many of which are highly toxic to cells (3). In addition, reactive aldehydes generated by lipid peroxidation can attack other cellular targets, such as proteins and DNA, thereby propagating the initial damage in cellular membranes to other macromolecules. Because lipid hydroperoxides formed in membranes are an important component of ROS generation in vivo, their detoxification appears to ...

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Section: Discussionmentioning

confidence: 99%

Transgenic Mice Overexpressing Glutathione Peroxidase 4 Are Protected against Oxidative Stress-induced Apoptosis

Ran

Liang

et al. 2004

Journal of Biological Chemistry

229

192

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“…The primers were a sense primer (5Ј-AGCT-TCTCTGTCTACCCTCCT-3Ј) and an antisense primer (5Ј-CGGGGT-CAGAGTTTCATCCC-3Ј), corresponding to the sequence from nucleotide 38 to 58 and that from nucleotide 548 to 568, respectively. Polymerase chain reaction was carried out in a MiniCycler (MJ Research, Inc., Watertown, MA) with Pfu DNA polymerase under the same conditions as described previously (15) except that the temperature of the annealing step was 55°C. After isopropyl alcohol precipitation, the amplified cDNA was ligated into the HincII site of pGEM3Zf(ϩ).…”

Section: Construction Of An Expression Plasmid For Production Of a Fumentioning

confidence: 99%

Identification of S100b Protein as Copper-binding Protein and Its Suppression of Copper-induced Cell Damage

Nishikawa

Lee

Shiraishi

et al. 1997

Journal of Biological Chemistry

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We have isolated from bovine brain a protein with a high capacity to inhibit the copper ion-catalyzed oxidation of L-ascorbate and identified it as S100b protein, an EF-hand calcium-binding protein, by sequencing its proteolytic peptides. Copper binding studies showed that this protein has four copper-binding sites per dimeric protein molecule with a dissociation constant of 0.46 M and that in the presence of L-ascorbate, copper ions bind to a total of six binding sites with a great increase in affinity. Furthermore, we examined whether S100b protein can prevent copper-induced cell damage. Bovine S100b protein was found to suppress dose-dependently the hemolysis of mouse erythrocytes induced by CuCl 2 . We transformed Escherichia coli cells with pGEX-5X-3 vector containing a cDNA for rat S100b protein, so that this protein could be expressed as a fusion protein with glutathione S-transferase. The transformed cells were demonstrated to be markedly resistant to a treatment with CuCl 2 plus H 2 O 2 as compared with the control cells expressing glutathione S-transferase alone. These results indicate that S100b protein does suppress oxidative cell damage by sequestering copper ions.

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“…The role of PHGPx in the cellular defense against oxidative stress has been widely investigated in several cell models in the past decade (12)(13)(14)(15)(16). Overexpression of a human PHGPx cDNA in guinea pig cells protected host cells from exogenous phosphatidylcholine hydroperoxide-mediated injury (13).…”

Section: Introductionmentioning

confidence: 99%

“…Overexpression of a human PHGPx cDNA in guinea pig cells protected host cells from exogenous phosphatidylcholine hydroperoxide-mediated injury (13). Moreover, yeast and tobacco cells expressing the tomato PHGPx gene (LePHGPx) were also resistant to cell death induced by oxidative stress and Bax (14), and the expression of RsPHGPx in a yeast PHGPx-deletion mutant significantly rescued the mutant sensitivity to oxidation-sensitive linelenic acid, just as the yeast PHGPx3 gene (ScPHGPx3) did (12 In this study, RsPHGPx was purified and added into cultured mouse 3T3 fibroblasts to examine its ability to protect cells against hydroperoxide-mediated injury.…”

Section: Introductionmentioning

confidence: 99%

Radish phospholipid hydroperoxide glutathione peroxidase provides protection against hydroperoxide-mediated injury in mouse 3T3 fibroblasts

Tian

Liu²,

Duan³

et al. 2009

BMB Reports

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Expression of Human Phospholipid Hydroperoxide Glutathione Peroxidase Gene for Protection of Host Cells from Lipid Hydroperoxide-Mediated Injury

Cited by 72 publications

References 0 publications

Transgenic Mice Overexpressing Glutathione Peroxidase 4 Are Protected against Oxidative Stress-induced Apoptosis

Transgenic Mice Overexpressing Glutathione Peroxidase 4 Are Protected against Oxidative Stress-induced Apoptosis

Identification of S100b Protein as Copper-binding Protein and Its Suppression of Copper-induced Cell Damage

Radish phospholipid hydroperoxide glutathione peroxidase provides protection against hydroperoxide-mediated injury in mouse 3T3 fibroblasts

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