Structural and Biochemical Characterization of an Active Arylamine N-Acetyltransferase Possessing a Non-canonical Cys-His-Glu Catalytic Triad - Archive ouverte HAL Access content directly
Journal Articles Journal of Biological Chemistry Year : 2013

Structural and Biochemical Characterization of an Active Arylamine N-Acetyltransferase Possessing a Non-canonical Cys-His-Glu Catalytic Triad

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Abstract

Arylamine N-acetyltransferases (NATs), a class of xenobiotic-metabolizing enzymes, catalyze the acetylation of aromatic amine compounds through a strictly conserved Cys-His-Asp catalytic triad. Each residue is essential for catalysis in both prokaryotic and eukaryotic NATs. Indeed, in (HUMAN)NAT2 variants, mutation of the Asp residue to Asn, Gln, or Glu dramatically impairs enzyme activity. However, a putative atypical NAT harboring a catalytic triad Glu residue was recently identified in Bacillus cereus ((BACCR)NAT3) but has not yet been characterized. We report here the crystal structure and functional characterization of this atypical NAT. The overall fold of (BACCR)NAT3 and the geometry of its Cys-His-Glu catalytic triad are similar to those present in functional NATs. Importantly, the enzyme was found to be active and to acetylate prototypic arylamine NAT substrates. In contrast to (HUMAN) NAT2, the presence of a Glu or Asp in the triad of (BACCR)NAT3 did not significantly affect enzyme structure or function. Computational analysis identified differences in residue packing and steric constraints in the active site of (BACCR)NAT3 that allow it to accommodate a Cys-His-Glu triad. These findings overturn the conventional view, demonstrating that the catalytic triad of this family of acetyltransferases is plastic. Moreover, they highlight the need for further study of the evolutionary history of NATs and the functional significance of the predominant Cys-His-Asp triad in both prokaryotic and eukaryotic forms. Background: Catalytic activity of prokaryotic and eukaryotic arylamine N-acetyltransferases (NATs) relies on a strictly conserved catalytic Cys-His-Asp triad. Results: Structural and biochemical studies identified a functional NAT with a Cys-His-Glu catalytic triad. Conclusion: The catalytic triad of these acetyltransferases is more plastic than previously believed. Significance: (BACCR)NAT3 represents the first known functional acetyltransferase with a non-canonical Cys-His-Glu catalytic triad.
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pasteur-03525397 , version 1 (13-01-2022)

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Attribution - CC BY 4.0

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Xavier Kubiak, Inès Li de La Sierra-Gallay, Alain F. Chaffotte, Benjamin Pluvinage, Patrick Weber, et al.. Structural and Biochemical Characterization of an Active Arylamine N-Acetyltransferase Possessing a Non-canonical Cys-His-Glu Catalytic Triad. Journal of Biological Chemistry, 2013, 288 (31), pp.22493 - 22505. ⟨10.1074/jbc.m113.468595⟩. ⟨pasteur-03525397⟩
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