Time-Resolved Crystallography of the Reaction Intermediate of Nitrile Hydratase: Revealing a Role for the Cysteinesulfenic Acid Ligand as a Catalytic Nucleophile

Angewandte Chemie
Yasuaki YamanakaMasafumi Odaka

Abstract

The reaction mechanism of nitrile hydratase (NHase) was investigated using time-resolved crystallography of the mutant NHase, in which βArg56, strictly conserved and hydrogen bonded to the two post-translationally oxidized cysteine ligands, was replaced by lysine, and pivalonitrile was the substrate. The crystal structures of the reaction intermediates were determined at high resolution (1.2-1.3 Å). In combination with FTIR analyses of NHase following hydration in H2 (18) O, we propose that the metal-coordinated substrate is nucleophilically attacked by the O(SO(-) ) atom of αCys114-SO(-) , followed by nucleophilic attack of the S(SO(-) ) atom by a βArg56-activated water molecule to release the product amide and regenerate αCys114-SO(-) .

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Citations

Mar 24, 2016·The Journal of Physical Chemistry. B·Megumi KayanumaYasuteru Shigeta
May 12, 2020·Frontiers in Bioengineering and Biotechnology·Zhongyi ChengZhemin Zhou
Aug 20, 2020·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·K P Wasantha LankathilakaRichard C Holz
Feb 13, 2016·The Journal of Biological Chemistry·Micah T NelpVahe Bandarian
Jun 16, 2019·Molecular Biology Reports·K SupreethaS Kiran
Dec 30, 2016·Journal of the American Chemical Society·Gloria Villar-AcevedoJulie A Kovacs
Oct 14, 2021·Dalton Transactions : an International Journal of Inorganic Chemistry·David S Glueck
Nov 30, 2021·Biochemistry·Wasantha Lankathilaka Karunagala PathiranageBrian Bennett

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