The role of glutathione in disulphide bond formation and endoplasmic‐reticulum‐generated oxidative stress

Chakravarthi, Seema; Jessop, Catherine E.; Bulleid, Neil J.

doi:10.1038/sj.embor.7400645

Cited by 385 publications

(306 citation statements)

References 32 publications

(51 reference statements)

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“…Third, GSH may play an important role in oxidative protein folding that is disrupted as a result of 17-AAG blockade of Hsp90 chaperoning functions. Recent studies have shown that GSH is required to regulate formation of certain native disulfide bonds, thereby contributing to correct folding of substrate proteins (47). When this process is disrupted due to endoplasmic reticulum stress, which may occur as a result of ansamycin treatment, reactive oxygen species may be released from the endoplasmic reticulum.…”

Section: Discussionmentioning

confidence: 99%

Up-regulation of Heat Shock Protein 27 Induces Resistance to 17-Allylamino-Demethoxygeldanamycin through a Glutathione-Mediated Mechanism

McCollum¹,

TenEyck

Sauer³

et al. 2006

Cancer Research

151

152

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Section: Discussionmentioning

confidence: 99%

Up-regulation of Heat Shock Protein 27 Induces Resistance to 17-Allylamino-Demethoxygeldanamycin through a Glutathione-Mediated Mechanism

McCollum¹,

TenEyck

Sauer³

et al. 2006

Cancer Research

151

152

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“…Additionally, we note that reported estimates of HeLa cell volume (2;600 μm 3 ) (23) and typical concentrations of cellular reduced glutathione (ca. 5 mM) (24) suggest that PL is present at quantities greatly in excess of cellular glutathione under our assay conditions (1,000 cells per well, 50 μL per well). As such, direct conjugation of PL with glutathione at C3 is a plausible explanation for the observed decrease in total glutathione.…”

Section: Discussionmentioning

confidence: 99%

Synthesis, cellular evaluation, and mechanism of action of piperlongumine analogs

Adams

Dai

Pellegrino

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

208

221

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Piperlongumine is a naturally occurring small molecule recently identified to be toxic selectively to cancer cells in vitro and in vivo. This compound was found to elevate cellular levels of reactive oxygen species (ROS) selectively in cancer cell lines. The synthesis of 80 piperlongumine analogs has revealed structural modifications that retain, enhance, and ablate key piperlongumine-associated effects on cells, including elevation of ROS, cancer cell death, and selectivity for cancer cells over nontransformed cell types. Structure/activity relationships suggest that the electrophilicity of the C2-C3 olefin is critical for the observed effects on cells. Furthermore, we show that analogs lacking a reactive C7-C8 olefin can elevate ROS to levels observed with piperlongumine but show markedly reduced cell death, suggesting that ROS-independent mechanisms, including cellular cross-linking events, may also contribute to piperlongumine's induction of apoptosis. In particular, we have identified irreversible protein glutathionylation as a process associated with cellular toxicity. We propose a mechanism of action for piperlongumine that may be relevant to other small molecules having two sites of reactivity, one with greater and the other with lesser electrophilicity.

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“…Heterologous protein production often results in the accumulation of reactive oxygen species (ROS) due to an increased demand for protein folding in the ER (23). The ROS accumulation in Y1EK was set as 1 relative fluorescence unit (RFU) (Fig.…”

Section: Figmentioning

confidence: 99%

Moderate Expression of SEC16 Increases Protein Secretion by Saccharomyces cerevisiae

Bao

Huang

Petranović

et al. 2017

Appl Environ Microbiol

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The yeast Saccharomyces cerevisiae is widely used to produce biopharmaceutical proteins. However, the limited capacity of the secretory pathway may reduce its productivity. Here, we increased the secretion of a heterologous ␣-amylase, a model protein used for studying the protein secretory pathway in yeast, by moderately overexpressing SEC16, which is involved in protein translocation from the endoplasmic reticulum to the Golgi apparatus. The moderate overexpression of SEC16 increased ␣-amylase secretion by generating more endoplasmic reticulum exit sites. The production of reactive oxygen species resulting from the heterologous ␣-amylase production was reduced. A genome-wide expression analysis indicated decreased endoplasmic reticulum stress in the strain that moderately overexpressed SEC16, which was consistent with a decreased volume of the endoplasmic reticulum. Additionally, fewer mitochondria were observed. Finally, the moderate overexpression of SEC16 was shown to improve the secretion of two other recombinant proteins, Trichoderma reesei endoglucanase I and Rhizopus oryzae glucan-1,4-␣-glucosidase, indicating that this mechanism is of general relevance.IMPORTANCE There is an increasing demand for recombinant proteins to be used as enzymes and pharmaceuticals. The yeast Saccharomyces cerevisiae is a cell factory that is widely used to produce recombinant proteins. Our study revealed that moderate overexpression of SEC16 increased recombinant protein secretion in S. cerevisiae. This new strategy can be combined with other targets to engineer cell factories to efficiently produce protein in the future. KEYWORDS protein secretion, mitochondria, ERES, ROS, SEC16T he yeast Saccharomyces cerevisiae is a widely used cell factory for the production of recombinant proteins (1). The advantages of S. cerevisiae are fast growth, easy cultivation, and ability to perform posttranslational modifications and secrete proteins to the extracellular medium, which facilitates the purification of the protein (2, 3). However, S. cerevisiae naturally secretes only a few proteins, such as aspartyl protease, invertase, and ␣-factor pheromone (4, 5); therefore, it has evolved a secretory pathway with a relatively low capacity. Many different strategies have been used to engineer this pathway with the objective of improving heterologous protein production (6-8). When expressing a protein that has a secretory signal peptide, the protein is cotranslationally translocated into the endoplasmic reticulum (ER) lumen, where disulfide bond formation occurs with the initial glycosylation (9). The secretion of recombinant proteins is affected by their signal peptide sequences (10). A synthetic leader sequence increases human insulin precursor production, whereas the use of the ␣-factor leader sequence increased ␣-amylase production (7). Excessive mannose glycosylation of recombinant proteins can reduce the efficiency of protein secretion, and it was recently found that

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The role of glutathione in disulphide bond formation and endoplasmic‐reticulum‐generated oxidative stress

Cited by 385 publications

References 32 publications

Up-regulation of Heat Shock Protein 27 Induces Resistance to 17-Allylamino-Demethoxygeldanamycin through a Glutathione-Mediated Mechanism

Up-regulation of Heat Shock Protein 27 Induces Resistance to 17-Allylamino-Demethoxygeldanamycin through a Glutathione-Mediated Mechanism

Synthesis, cellular evaluation, and mechanism of action of piperlongumine analogs

Moderate Expression of SEC16 Increases Protein Secretion by Saccharomyces cerevisiae

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