Discovery and Substrate Specificity Engineering of Nucleotide Halogenases

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Jun 19

PMID 38898020

Authors

Jie Ni

Jingyuan Zhuang

Yiming Shi

Ying-Chih Chiang

Gui-Juan Cheng

Affiliations

Soon will be listed here.

Abstract

C2'-halogenation has been recognized as an essential modification to enhance the drug-like properties of nucleotide analogs. The direct C2'-halogenation of the nucleotide 2'-deoxyadenosine-5'-monophosphate (dAMP) has recently been achieved using the Fe(II)/α-ketoglutarate-dependent nucleotide halogenase AdaV. However, the limited substrate scope of this enzyme hampers its broader applications. In this study, we report two halogenases capable of halogenating 2'-deoxyguanosine monophosphate (dGMP), thereby expanding the family of nucleotide halogenases. Computational studies reveal that nucleotide specificity is regulated by the binding pose of the phosphate group. Based on these findings, we successfully engineered the substrate specificity of these halogenases by mutating second-sphere residues. This work expands the toolbox of nucleotide halogenases and provides insights into the regulation mechanism of nucleotide specificity.

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