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- Title
Discovery and substrate specificity engineering of nucleotide halogenases.
- Authors
Ni, Jie; Zhuang, Jingyuan; Shi, Yiming; Chiang, Ying-Chih; Cheng, Gui-Juan
- Abstract
C2′-halogenation has been recognized as an essential modification to enhance the drug-like properties of nucleotide analogs. The direct C2ʹ-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 2ʹ-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. The direct C2ʹ halogenation of the nucleotide 2′-deoxyadenosine-5′-monophosphate (dAMP) was achieved using the Fe(II)/α-ketoglutarate dependent nucleotide halogenase AdaV, but the limited substrate scope of this enzyme hinders its application. Here, the authors discover two halogenases capable of halogenating 2ʹ-deoxyguanosine monophosphate (dGMP) and reveal that nucleotide specificity is regulated by the binding pose of the phosphate group.
- Subjects
HALOGENASES; BIOCHEMICAL substrates; HALOGENATION; ENGINEERING; ENZYMES
- Publication
Nature Communications, 2024, Vol 15, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-024-49147-7