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Nit1 is a metabolite repair enzyme that hydrolyzes deaminated glutathione.

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Peracchi, Alessio Veiga da Cunha, Maria [UCL] Kuhara, Tomiko Ellens, Kenneth W Paczia, Nicole Bommer, Guido [UCL] Van Schaftingen, Emile [UCL]

The mammalian gene Nit1 (nitrilase-like protein 1) encodes a protein that is highly conserved in eukaryotes and is thought to act as a tumor suppressor. Despite being ∼35% sequence identical to ω-amidase (Nit2), the Nit1 protein does not hydrolyze efficiently α-ketoglutaramate (a known physiological substrate of Nit2), and its actual enzymatic function has so far remained a puzzle. In the present study, we demonstrate that both the mammalian Nit1 and its yeast ortholog are amidases highly active toward deaminated glutathione (dGSH; i.e., a form of glutathione in which the free amino group has been replaced by a carbonyl group). We further show that Nit1-KO mutants of both human and yeast cells accumulate dGSH and the same compound is excreted in large amounts in the urine of Nit1-KO mice. Finally, we show that several mammalian aminotransferases (transaminases), both cytosolic and mitochondrial, can form dGSH via a common (if slow) side-reaction and provide indirect evidence that transaminases are mainly responsible for dGSH formation in cultured mammalian cells. Altogether, these findings delineate a typical instance of metabolite repair, whereby the promiscuous activity of some abundant enzymes of primary metabolism leads to the formation of a useless and potentially harmful compound, which needs a suitable "repair enzyme" to be destroyed or reconverted into a useful metabolite. The need for a dGSH repair reaction does not appear to be limited to eukaryotes: We demonstrate that Nit1 homologs acting as excellent dGSH amidases also occur in Escherichia coli and other glutathione-producing bacteria.

Document type Article de périodique (Journal article) – Article de recherche
Publication date 2017
Language Anglais
Journal information "Proceedings of the National academy of sciences of the United States of America" - Vol. 114, no.16, p. E3233-E3242 (2017)
Peer reviewed yes
Publisher National academy of sciences
issn 0027-8424
e-issn 1091-6490
Publication status Publié
Affiliation UCL - SSS/DDUV/BCHM - Biochimie-Recherche métabolique
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Bibliographic reference Peracchi, Alessio ; Veiga da Cunha, Maria ; Kuhara, Tomiko ; Ellens, Kenneth W ; Paczia, Nicole ; et. al. Nit1 is a metabolite repair enzyme that hydrolyzes deaminated glutathione.. In: Proceedings of the National academy of sciences of the United States of America, Vol. 114, no.16, p. E3233-E3242 (2017)
Permanent URL http://hdl.handle.net/2078.1/185732