Identification and Characterization of Pyrimidine Nucleoside 2′-Hydroxylase

IF 11.3 1区化学 Q1 CHEMISTRY, PHYSICAL

ACS Catalysis Pub Date : 2025-02-13 DOI:10.1021/acscatal.4c07764

Ferdinand Genz, Florian Friedrich, Christoph Lönarz, Oliver Einsle, Manfred Jung, Michael Müller, Nico D. Fessner

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Abstract

Functionalization of nucleosides at the 2′-position has become an important modification for therapeutic purposes to tailor pharmacological properties. The chemical synthesis of these molecules is challenging, and recent studies have explored bottom-up strategies with enzymes of the nucleoside salvage pathway. More than 50 years ago, a pyrimidine nucleoside 2′-hydroxylase (PDN2′H) activity had been described in fungal species extracts. However, the corresponding protein sequences were never reported and the protein characterization remained incomplete. This study describes the identification and characterization of PDN2′H from Neurospora crassa, which naturally hydroxylates thymidine at the α-2′-position as was now verified by NMR spectroscopy. Site-directed mutagenesis and biochemical assays indicated the protein to be an α-ketoglutarate-/Fe(II)-dependent dioxygenase. Furthermore, the substrate scope, phylogeny, and thermostability of NcPDN2′H were determined and its enzymatic mechanism was elucidated by resolving its X-ray protein structure cocrystallized with thymidine. NcPDN2′H is a long sought-after and important nucleoside-modifying addition to the biocatalytic portfolio.

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来源期刊

ACS Catalysis CHEMISTRY, PHYSICAL-

CiteScore

20.80

自引率

6.20%

发文量

1253

审稿时长

1.5 months

期刊介绍： ACS Catalysis is an esteemed journal that publishes original research in the fields of heterogeneous catalysis, molecular catalysis, and biocatalysis. It offers broad coverage across diverse areas such as life sciences, organometallics and synthesis, photochemistry and electrochemistry, drug discovery and synthesis, materials science, environmental protection, polymer discovery and synthesis, and energy and fuels. The scope of the journal is to showcase innovative work in various aspects of catalysis. This includes new reactions and novel synthetic approaches utilizing known catalysts, the discovery or modification of new catalysts, elucidation of catalytic mechanisms through cutting-edge investigations, practical enhancements of existing processes, as well as conceptual advances in the field. Contributions to ACS Catalysis can encompass both experimental and theoretical research focused on catalytic molecules, macromolecules, and materials that exhibit catalytic turnover.