期刊
XENOBIOTICA
卷 47, 期 2, 页码 164-175出版社
TAYLOR & FRANCIS LTD
DOI: 10.3109/00498254.2016.1166533
关键词
Acetaminophen toxicity; glutathione deficiency; gamma-glutamyl cycle; protein arylation
资金
- Southampton Hospital Charity, Southampton General Hospital, UK [1051543, 0182]
- National Institute of Health (NIH) [R15GM086833]
- Texas Woman's University Research Enhancement Grant
1. Metabolic acidosis due to accumulation of L-5-oxoproline is a rare, poorly understood, disorder associated with acetaminophen treatment in malnourished patients with chronic morbidity. L-5-Oxoprolinuria signals abnormal functioning of the gamma-glutamyl cycle, which recycles and synthesises glutathione. Inhibition of glutathione synthetase (GS) by N-acetylp- benzoquinone imine (NAPQI) could contribute to 5-oxoprolinuric acidosis in such patients. We investigated the interaction of NAPQI with GS in vitro. 2. Peptide mapping of co-incubated NAPQI and GS using mass spectrometry demonstrated binding of NAPQI with cysteine-422 of GS, which is known to be essential for GS activity. Computational docking shows that NAPQI is properly positioned for covalent bonding with cysteine-422 via Michael addition and hence supports adduct formation. 3. Co-incubation of 0.77 mu M of GS with NAPQI (25-400 mu M) decreased enzyme activity by 1689%. Inhibition correlated strongly with the concentration of NAPQI and was irreversible. 4. NAPQI binds covalently to GS causing irreversible enzyme inhibition in vitro. This is an important novel biochemical observation. It is the first indication that NAPQI may inhibit glutathione synthesis, which is pivotal in NAPQI detoxification. Further studies are required to investigate its biological significance and its role in 5-oxoprolinuric acidosis.
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