Production and purification of homogenous recombinant human selenoproteins reveals a unique codon skipping event in E. coli and GPX4-specific affinity to bromosulfophthalein

Cheng, Qing; Roveri, Antonella; Cozza, Giorgio; Bordin, Luciana; Rohn, Isabelle; Schwerdtle, Tanja; Kipp, Anna P.; Ursini, Fulvio; Maiorino, Matilde; Miotto, Giovanni; Arnér, Elias S.J.

doi:10.1016/j.redox.2021.102070

Cited by 17 publications

(24 citation statements)

References 47 publications

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“…GPX4 is a selenoprotein that catalyzes glutathione to convert lipid peroxides into lipid alcohols [ 52 ], effectively removing the accumulation of lipid peroxides in cells and providing antioxidant defense against ferroptosis [ 53 ]. GSH is the most crucial cofactor of GPX4, and the expression level of GSH will directly affect the activity of GPX4.…”

Section: New Approaches To Metal Ions-induced Cell Death and Their An...mentioning

confidence: 99%

“…Producing reactive oxygen by Fenton reaction Fe 3 O 4 /Gd 2 O 3 @CDDP@LF/RGD2 Fe 3 O 4 , CDDP [49] Gox@MOF(Fe)@CCM Fe 2+ , Fe 3+ , GOx [50] AuFCSP MOFs Fe 2+ , Fe 3+ , Au NPs [51] Inhibiting the activity of GXP4 RSL3@COF-Fc Fc, RSL3 [56] HMPB/ ML210@TA-BLM-Fe 3+ Fe 2+ , Fe 3+ , ML210 [57] Consuming intracellular GSH CA-OH-Fe 3+ /Gd 3+ @P-SS-D Fe 2+ , Fe 3+ , CA [58] siMCT4-PAMAM-PEG-TK-Fc@DEM Fc, DEM, siMCT4 [59] FPBC@SN Fe 2+ , Fe 3+ , SRF, NLG919 [60] Inhibiting the activity of System Xc-Ce6-Erastin Erastin [66] SSZ-Fe 2+ @DSSD Fe 2+ , SSZ [67] MIL-101(Fe)@SRF Fe 2+ , Fe 3+ , SRF [68] Delivering exogenous lipid peroxides IO-LAHP IO NPs, LAHP [71] RSL3@mPEG-PLys-AA RSL3, AA [72] Lecithin@FAC Fe 2+ , Fe 3+ , Lecithin [73] H 2 O 2 are crucial to the above process. GPX4 is a selenoprotein that catalyzes glutathione to convert lipid peroxides into lipid alcohols [52], effectively removing the accumulation of lipid peroxides in cells and providing antioxidant defense against ferroptosis [53]. GSH is the most crucial cofactor of GPX4, and the expression level of GSH will directly affect the activity of GPX4.…”

Section: Nanomaterials Example Ferroptosis Components Referencesmentioning

confidence: 99%

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New anti-cancer explorations based on metal ions

et al. 2022

J Nanobiotechnol

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Due to the urgent demand for more anti-cancer methods, the new applications of metal ions in cancer have attracted increasing attention. Especially the three kinds of the new mode of cell death, including ferroptosis, calcicoptosis, and cuproptosis, are of great concern. Meanwhile, many metal ions have been found to induce cell death through different approaches, such as interfering with osmotic pressure, triggering biocatalysis, activating immune pathways, and generating the prooxidant effect. Therefore, varieties of new strategies based on the above approaches have been studied and applied for anti-cancer applications. Moreover, many contrast agents based on metal ions have gradually become the core components of the bioimaging technologies, such as MRI, CT, and fluorescence imaging, which exhibit guiding significance for cancer diagnosis. Besides, the new nano-theranostic platforms based on metal ions have experimentally shown efficient response to endogenous and exogenous stimuli, which realizes simultaneous cancer therapy and diagnosis through a more controlled nano-system. However, most metal-based agents have still been in the early stages, and controlled clinical trials are necessary to confirm or not the current expectations. This article will focus on these new explorations based on metal ions, hoping to provide some theoretical support for more anti-cancer ideas.

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Section: New Approaches To Metal Ions-induced Cell Death and Their An...mentioning

confidence: 99%

Section: Nanomaterials Example Ferroptosis Components Referencesmentioning

confidence: 99%

New anti-cancer explorations based on metal ions

et al. 2022

J Nanobiotechnol

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“…Possible alternative in-frame decoding mechanisms following amino acid deprivation are codon reassignment and translational bypass. These processes result in the incorporation of a different amino acid than is encoded for in the mRNA and the interruption of translation followed by downstream continuation 9 , 10 , 16 , 17 (Fig. 1a ).…”

Section: Codon Reassignment By Tryptophan Depletionmentioning

confidence: 99%

Tryptophan depletion results in tryptophan-to-phenylalanine substitutants

Pataskar

Champagne

Nagel

et al. 2022

Nature

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Activated T cells secrete interferon-γ, which triggers intracellular tryptophan shortage by upregulating the indoleamine 2,3-dioxygenase 1 (IDO1) enzyme1–4. Here we show that despite tryptophan depletion, in-frame protein synthesis continues across tryptophan codons. We identified tryptophan-to-phenylalanine codon reassignment (W>F) as the major event facilitating this process, and pinpointed tryptophanyl-tRNA synthetase (WARS1) as its source. We call these W>F peptides ‘substitutants’ to distinguish them from genetically encoded mutants. Using large-scale proteomics analyses, we demonstrate W>F substitutants to be highly abundant in multiple cancer types. W>F substitutants were enriched in tumours relative to matching adjacent normal tissues, and were associated with increased IDO1 expression, oncogenic signalling and the tumour-immune microenvironment. Functionally, W>F substitutants can impair protein activity, but also expand the landscape of antigens presented at the cell surface to activate T cell responses. Thus, substitutants are generated by an alternative decoding mechanism with potential effects on gene function and tumour immunoreactivity.

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“…Translational frameshifting has, to our knowledge, not been explored in the context of canonical selenoproteins. There is, however, a known instant where a synthetic UAG/Sec codon in GPX4 recombinantly expressed in an RF1 -mutant E. coli was skipped in a +3 frameshifting event [ 55 ]. This may indicate that ribosomes apparently try to avoid termination in a codon context not optimized for termination [ 50 ].…”

Section: Discussionmentioning

confidence: 99%

High-Resolution Ribosome Profiling Reveals Gene-Specific Details of UGA Re-Coding in Selenoprotein Biosynthesis

et al. 2022

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Co-translational incorporation of selenocysteine (Sec) into selenoproteins occurs at UGA codons in a process in which translational elongation competes with translational termination. Selenocysteine insertion sequence-binding protein 2 (SECISBP2) greatly enhances Sec incorporation into selenoproteins by interacting with the mRNA, ribosome, and elongation factor Sec (EFSEC). Ribosomal profiling allows to study the process of UGA re-coding in the physiological context of the cell and at the same time for all individual selenoproteins expressed in that cell. Using HAP1 cells expressing a mutant SECISBP2, we show here that high-resolution ribosomal profiling can be used to assess read-through efficiency at the UGA in all selenoproteins, including those with Sec close to the C-terminus. Analysis of ribosomes with UGA either at the A-site or the P-site revealed, in a transcript-specific manner, that SECISBP2 helps to recruit tRNASec and stabilize the mRNA. We propose to assess the effect of any perturbation of UGA read-through by determining the proportion of ribosomes carrying UGA in the P-site, pUGA. An additional, new observation is frameshifting that occurred 3′ of the UGA/Sec codon in SELENOF and SELENOW in SECISBP2-mutant HAP1 cells, a finding corroborated by reanalysis of neuron-specific Secisbp2R543Q-mutant brains.

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Production and purification of homogenous recombinant human selenoproteins reveals a unique codon skipping event in E. coli and GPX4-specific affinity to bromosulfophthalein

Cited by 17 publications

References 47 publications

New anti-cancer explorations based on metal ions

New anti-cancer explorations based on metal ions

Tryptophan depletion results in tryptophan-to-phenylalanine substitutants

High-Resolution Ribosome Profiling Reveals Gene-Specific Details of UGA Re-Coding in Selenoprotein Biosynthesis

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