Impact of residues remote from the catalytic centre on enzyme catalysis of copper nitrite reductase

Nicole Leferink, Svetlana V. Antonyuk, Joseline A. Houwman, Nigel S. Scrutton, Robert R. Eady, S. Samar Hasnain

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Enzyme mechanisms are often probed by structure-informed point mutations and measurement of their effects on enzymatic properties to test mechanistic hypotheses. In many cases, the challenge is to report on complex, often inter-linked elements of catalysis. Evidence for long-range effects on enzyme mechanism resulting from mutations remains sparse, limiting the design/redesign of synthetic catalysts in a predictable way. Here we show that improving the accessibility of the active site pocket of copper nitrite reductase by mutation of a surface-exposed phenylalanine residue (Phe306), located 12 €‰Å away from the catalytic site type-2 Cu (T2Cu), profoundly affects intra-molecular electron transfer, substrate-binding and catalytic activity. Structures and kinetic studies provide an explanation for the lower affinity for the substrate and the alteration of the rate-limiting step in the reaction. Our results demonstrate that distant residues remote from the active site can have marked effects on enzyme catalysis, by driving mechanistic change through relatively minor structural perturbations. © 2014 Macmillan Publishers Limited. All rights reserved.
    Original languageEnglish
    Article number4395
    JournalNature Communications
    Volume5
    DOIs
    Publication statusPublished - 15 Jul 2014

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