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image of Structural Insights into Ligand-Induced Conformational Changes in Adenine Phosphoribosyl Transferase from Fusobacterium nucleatum

Abstract

Background

Adenine phosphoribosyltransferase (APRT) is an enzyme that facilitates adenosine monophosphate (AMP) biosynthesis by transferring a phosphoribosyl group to adenine using phosphoribosyl pyrophosphate as a donor. While the human enzyme is well characterized, structural insights into bacterial APRTs remain limited. is associated with periodontal disease, yet its APRT enzyme (APRT) has not been structurally investigated.

Objective

This study aimed to examine the crystal structure of APRT and ligand-induced conformational changes to understand its enzymatic and substrate recognition mechanisms.

Methods

The APRT protein was heterologously expressed in , followed by initial purification using nickel-charged affinity resin chromatography and further purification through size-exclusion chromatography. The APRT structure was resolved using X-ray crystallography and compared with that of APRT (APRT), exhibiting the highest amino acid sequence similarity among bacterial APRT structures.

Results

AMP and phosphate (PO) were observed in the active site of APRT. Significant differences in ligand positioning were observed between the AMP-PO-bound structures of APRT and APRT. Structural shifts induced by AMP-PO binding were detected. The Arg78 and Lys82 residues from the alternate subunit occupied the PO site in the absence of ligands, but they interacted with PO upon AMP-PO binding. Structural comparison of the AMP-PO-bound APRT with that of the adenine-bound APRT highlighted variations in the adenine-binding site and associated structural changes.

Discussion

Structural comparison of the AMP-PO-bound APRT with that of the adenine-bound APRT highlighted variations in the adenine-binding site and the associated structural changes.

Conclusion

The AMP-PO-bound APRT exhibited distinct ligand-binding modes despite sharing a high sequence similarity with APRT. The structures demonstrated ligand movement during bacterial APRT reactions.

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2026-01-22
2026-02-16

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Structural Insights into Ligand-Induced Conformational Changes in Adenine Phosphoribosyl Transferase from Fusobacterium nucleatum
Bentham Science Publishers ; https://doi.org/10.2174/0109298665403166251021110505
/content/journals/ppl/10.2174/0109298665403166251021110505
/content/journals/ppl/10.2174/0109298665403166251021110505
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Purification, crystallization, and X-ray diffraction of adenine phosphoribosyltransferase APRT Electron density maps around the ligand-binding site of APRT. Atomic-level interactions between APRT (chain A) and the symmetry-related molecule (chain B). Comparative interaction analysis of AMP-PO binding residues in APRT and APRT. Supplementary material is available on the publisher’s website along with the published article.

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  • Article Type:     Research Article
Keywords: crystal structure ; Fusobacterium nucleatum ; X-ray crystallography ; FnAPRT ; bacteria APRT ; conformation change ; Adenine phosphoribosyltransferase

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