8VOU

Human glutathione transferase M1-1 in complex with the adduct between glutathione and nitrooleic acid

This is a non-PDB format compatible entry.

Summary for 8VOU

• Entry DOI: 10.2210/pdb8vou/pdb
• Descriptor: Glutathione S-transferase Mu 1, L-gamma-glutamyl-S-[(8R,9S)-1-carboxy-9-nitroheptadecan-8-yl]-L-cysteinylglycine, GLUTATHIONE, ... (4 entities in total)
• Functional Keywords: complex, nitroalkene fatty acid, glutathione, transferase, transferase-product complex, transferase/product
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 4
• Total formula weight: 105833.82

Authors

Larrieux, N.,Steglich, M.,Dalla Rizza, J.,Buschiazzo, A.,Turell, L. (deposition date: 2024-01-16, release date: 2025-03-12, Last modification date: 2025-04-09)

Primary citation

Steglich, M.,Larrieux, N.,Zeida, A.,Dalla Rizza, J.,Salvatore, S.R.,Bonilla, M.,Moller, M.N.,Buschiazzo, A.,Alvarez, B.,Schopfer, F.J.,Turell, L.
Human glutathione transferases catalyze the reaction between glutathione and nitrooleic acid.
J.Biol.Chem., 301:108362-108362, 2025

PubMed Abstract: Nitroalkene fatty acids (NO-FAs) are formed endogenously. They regulate cell signaling pathways and are being developed clinically to treat inflammatory diseases. NO-FAs are electrophilic and form thioether adducts with glutathione (GSH), which are exported from cells. Glutathione transferases (GSTs), a superfamily of enzymes, contribute to the cellular detoxification of hydrophobic electrophiles by catalyzing their conjugation to GSH. Herein, we evaluated the capacity of five human GSTs (M1-1, M2-2, M4-4, A4-4, and P1-1) to catalyze the reaction between nitrooleic acid (NO-OA) and GSH. The reaction was monitored by HPLC-ESI-MS/MS and catalytic activity was detected with hGSTs M1-1 and A4-4. Using stopped-flow spectrophotometry, a 1400 and 7500-fold increase in the apparent second-order rate constant was observed for hGST M1-1 and hGST A4-4, respectively, compared to the uncatalyzed reaction (pH 7.4, 25 °C), in part due to a higher availability of the thiolate. The crystal structure of hGST M1-1 in complex with the adduct was solved at 2.55 Å resolution, revealing that the ligand was bound within the reaction center, and establishing a foundation to build a model of hGST A4-4 in complex with the adduct. A larger number of interactions between the enzyme and the fatty acid were observed for hGST A4-4 compared to hGST M1-1, probably contributing to the increased catalysis. Altogether, these results show, for the first time, that hGSTs can catalyze the reaction between GSH and NO-FAs, likely affecting the signaling actions of these metabolites and expanding the repertoire of GST reactions.

PubMed: 40024478
DOI: 10.1016/j.jbc.2025.108362

Experimental method

X-RAY DIFFRACTION (2.55 Å)