Glutathione (gsh)

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Sourced in Germany, United States

Glutathione is a tripeptide composed of the amino acids glutamic acid, cysteine, and glycine. It is an important antioxidant and plays a crucial role in various cellular processes. Glutathione is involved in the detoxification of harmful substances, supports the immune system, and helps maintain cellular redox balance.

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Lab products found in correlation

7 protocols using glutathione (gsh)

Synthesis of GSH-MDI Adducts

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Reduced glutathione, GSH (CAS no. 70-18-8) and 4,4′-methylenebis(phenyl isocyanate) or MDI (CAS no. 101-68-8) were from Sigma-Aldrich (St. Louis, MO) and were of ≥98.0% purity. GSH was reacted with MDI under conditions that yield reaction products with the greatest capacity to carbamoylate human albumin, among the conditions tested in previously published studies.^{18 (link)} Briefly, 50 μL of 10% (w/v) MDI in acetone from JT Baker (Phillipsburg, NJ) was added dropwise with stirring to 25 mL of 10 mM GSH in 200 mM sodium phosphate, pH 7.4. The reaction mixture was rotated end-over-end for 2 h at 37 °C and then centrifuged at 10 000g, 0.2 μm filtered, aliquoted, and used immediately or snap frozen in LN₂ and stored at −80 °C until analysis could be performed.

Wisnewski A.V., Liu J, & Colangelo C.M. (2015). Glutathione Reaction Products with a Chemical Allergen, Methylene-diphenyl Diisocyanate, Stimulate Alternative Macrophage Activation and Eosinophilic Airway Inflammation. Chemical research in toxicology, 28(4), 729-737.

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Wheat Flour Characterization and Modification

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Four different German commercial wheat flours Type 550 were supplied by Rosenmühle (Landshut, Germany). Flour characteristics were analyzed according to methods of the AACC international (AACCi) and of the International Association for Cereal Science and Technology (ICC): the moisture (AACCi 44-01), protein content (AACCi 46-16, N × 5.7), ash (ICC 104/1), and falling number (AACCi 56-81). All specifications and characteristics of the four flours are summarized in Table 1. The use of various flours was intended to detect microstructural changes independent of the raw material. Ascorbic acid was supplied by Carl Roth GmbH + Co. KG (Karlsruhe, Germany), glutathione by VWR International GmbH (Darmstadt, Germany), transglutaminase (≥1000 units/g) and glucose oxidase (≥1100 units/g) by AB Enzymes GmbH (kindly provided by AB Enzymes GmbH, Darmstadt, Germany), potassium bromate (KBrO₃) by ThermoFisher GmbH (Karlsruhe, Germany), bromelain (≥3 units/mg protein) and d(+)glucose by Sigma-Aldrich Chemie GmbH (Steiheim, Germany), Rhodamine B by Merck KGaA (Darmstadt, Germany), rapeseed oil by Cargill Oil Packers bvba (Izegem Belgien) and shortening (ingredients: palm fat, coconut fat, rapeseed oil, water, emulsifier, NaCl, citric acid, aroma, carotin) by MeisterMarken (CSM Deutschland GmbH, Bingen, Germany).

Lucas I., Becker T, & Jekle M. (2018). Gluten Polymer Networks—A Microstructural Classification in Complex Systems. Polymers, 10(6), 617.

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Rapid Synthesis of Antioxidant-Enriched Sensors

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Magnesium nitrate hexahydrate (Mg(NO 3 ) 2 •6H 2 O), aluminum nitrate nonahydrate (Al(NO 3 ) 3 •6H 2 O), sodium hydroxide (NaOH) and lactic acid (85%) were purchased from Merck and used without purification. Copper(II) chloride dihydrate (CuCl 2 •2H 2 O), neocuproine (Nc), trolox, L(+)-ascorbic acid, eugenol, diosmin, gallic acid, glutathione (reduced), catechin hydrate, tannic acid, chlorogenic acid ( predominantly trans), sodium salicylate, sodium alginate (NaAlg), sodium chloride (NaCl) and ethanol were obtained from VWR International in analytical grade and used as received. For sensor preparation, cellulose filter paper (Whatman Grade 602H, VWR) and crayon were used. Ultrapure water was obtained from a Puranity TU3+ UV/UF system.

Szerlauth A., Szalma L., Muráth S., Sáringer S., Varga G., Li L, & Szilágyi I. (2022). Nanoclay-based sensor composites for the facile detection of molecular antioxidants. The Analyst, 147(7).

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Synthesis of Functionalized Organic Compounds

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1,3,5-Tribromobenzene (98%), trimethylsilyl-acetylene (98%), trimethylsilyl azide (94%), and diethylamine (99%) were purchased from Alfa Aesar (Ward Hill, MA, USA). Sodium nitrite (97%), phosphate buffered saline (PBS) tablets, and copper(II) chloride dihydrate were obtained from EMD Chemicals (Gibbstown, NJ, USA). Glutathione (98%) was purchased from AMRESCO (Solon, OH, USA). Low molecular weight chitosan (96% deacetylated) and copper(I) iodide (99.5%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Deionized water (18.2 MΩ·cm) was obtained from a Millipore Direct-Q water purification system (EMD Millipore, Billerica, MA, USA). Ultrahigh purity nitrogen and oxygen gases were supplied by Airgas (Denver, CO, USA). Bis(triphenylphosphine)-palladium(II) dichloride (98%) was purchased from TCI America (Portland, OR, USA). All materials were used as received without any further purification.

Neufeld M.J., Lutzke A., Tapia J.B, & Reynolds M.M. (2017). Metal−Organic Framework/Chitosan Hybrid Materials Promote Nitric Oxide Release from S‑Nitrosoglutathione in Aqueous Solution. ACS applied materials & interfaces, 9(6), 5139-5148.

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Oxidative Folding of KIIIA and S-m3-huwentoxin-IV

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Oxidative
folding of
KIIIA peptides was performed as described previously.^{29 (link)} Oxidative folding of S-m₃-huwentoxin-IV was
accomplished by glutathione-assisted folding at 25 °C overnight
under the following conditions: 15 μM reduced peptide in 0.1
M Tris-HCl (pH 8.0, Amresco, Solon, OH), 10% (v/v) isopropanol (Chem-Supply
Pty. Ltd.), and 5 mM reduced and 1 mM oxidized glutathione (Sigma-Aldrich).
The reaction was quenched by decreasing the pH to 2 using an ACN/TFA/H₂O mixture [1/1/1 (v/v/v)].

Peschel A., Cardoso F.C., Walker A.A., Durek T., Stone M.R., Braga Emidio N., Dawson P.E., Muttenthaler M, & King G.F. (2020). Two for the Price of One: Heterobivalent Ligand Design Targeting Two Binding Sites on Voltage-Gated Sodium Channels Slows Ligand Dissociation and Enhances Potency. Journal of Medicinal Chemistry, 63(21), 12773-12785.

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Cytosolic GPX4 Mutant Expression

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The compound selected from virtual screen was purchased from SPECS with purity of more than 95% (confirmed by SPECS with NMR and LC-MS; data are available on the website http://www.specs.net/). Reagents were purchased from Sigma–Aldrich unless otherwise noted. The pQE-30 bacterial expression plasmid of the U46C mutant of human cytosolic GPX4 (c-GPX4) was a generous gift from Professor Hartmut Kuhn (University Medicine Berlin-Charité, Germany). IPTG, PMSF, DTT, EDTA, and glutathione were from Amresco. Standard compounds for LC-MS/MS were purchased from Cayman Chemical. Calcium ionophore A23187 was obtained from J&K Chemical. Zileuton was purchased from Tocris Bioscience. The Symmetry C18 reverse-phase column (3.5 μm, 2.1 mm × 150 mm) was purchased from Waters Corp. HEK293T cells were received as a gift from Professor Jincai Luo (Peking University, China). The Dual-Glo Luciferase Assay System was from Promega. The pCDNA3.1 plasmid containing the gene encoding firefly luciferase was a generous gift from Professor Peng Chen (Peking University, China). Antibody against GPX4 was obtained from Abcam (no. ab125066). HT-1080 cells were the generous gift of Professor Chu Wang (Peking University, China).

Li C., Deng X., Xie X., Liu Y., Friedmann Angeli J.P, & Lai L. (2018). Activation of Glutathione Peroxidase 4 as a Novel Anti-inflammatory Strategy. Frontiers in Pharmacology, 9, 1120.

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Synthesis of GSH-MDI Conjugate

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Reduced glutathione, GSH (CAS # 70-18-8), and 4,4’-methylenebis(phenyl isocyanate) or MDI (CAS # 101-68-8) were from Sigma-Aldrich (St. Louis, MO) and were of ≥ 98.0% purity. GSH was reacted with MDI as previously described (Wisnewski, Liu et al. 2013 (link)). Briefly, 50 µl of 10% (w/v) MDI in acetone from JT Baker (Phillipsburg, NJ) was added dropwise with stirring to 25 ml of 10 mM GSH in 200 mM sodium phosphate, pH 7.4 (final acetone concentration 0.2% v/v). The reaction mixture was rotated end-over-end for 2 hours at 37°C, and then centrifuged at 10,000 g, and 0.2 µm filtered before use.

Wisnewski A.V., Liu J, & Nassar A. (2015). In vitro cleavage of diisocyanate-glutathione conjugates by human gamma-glutamyl transpeptidase-1. Xenobiotica; the fate of foreign compounds in biological systems, 46(8), 726-732.

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