Structural insight into EV-A71 3A protein and its interaction with a peptide inhibitor

Q: What is the clinical evidence from this study?

Population: Enterovirus A71 (EV-A71) infection / Hand, foot, and mouth disease (HFMD). Intervention: D-peptide inhibitor ER-DRI vs. Untreated apo-form 15N-labeled EV-A71 3A(1-57) protein (1:0 molar ratio). Primary outcome: Structural dynamics and intermolecular binding interactions between EV-A71 3A(1-57) protein and ER-DRI via NMR spectroscopy and molecular docking.

Q: What does this research mean for the field?

The D-peptide inhibitor ER-DRI prevents EV-A71 3A protein homodimerization by competitively binding to its dimeric α-helix interface. Novelty: ClaimNovelty.NOVEL_FINDING. Consensus alignment: ConsensusAlignment.NEUTRAL.

Yahui Liu; Panjing Lv; Wei Wang; Jiahai Zhang; Zhou Xi; Yang Qiu; Kun Cai; Haoran Zhang; Yuan Fang; Yan Li

doi:10.1016/j.virs.2023.09.004

September 25, 2023Virologica SinicaOpen Access

Structural insight into EV-A71 3A protein and its interaction with a peptide inhibitor

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Key result

D-peptide ER-DRI competitively blocks EV-A71 3A protein homodimerization at the dimeric α-helix interface.

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Key Points

To elucidate the solution structure of Enterovirus A71 non-structural protein 3A in its apo state and determine the molecular mechanism of its inhibition by the peptide ER-DRI.
Expressed and purified recombinant EV-A71 3A 1–57 using a 6×His-SUMO tag cleavage system to generate a soluble 63-amino-acid peptide construct.
Conducted 1H-15N heteronuclear NMR spectroscopy and 15N relaxation experiments on uniformly [13C, 15N]-labeled 3A 1–57 samples.
Predicted secondary structural elements from backbone chemical shift assignments using TALOS-N and evaluated inhibitor interactions.
Achieved backbone resonance assignment for 75% (42/56) of non-proline amino acid pairs in EV-A71 3A 1–57.
Identified rigid dihelical structures spanning residues 25–45 that mediate homodimerization, accompanied by a structurally disordered, flexible C-terminus across residues 46–63 in the apo state.
Revealed that the D-peptide ER-DRI inhibits dimeric EV-A71 3A through a competitive binding mechanism targeting the homodimerization interface.

Structured PICO

P

Population

Recombinant Enterovirus A71 3A(1-57) protein expressed in E. coli for biophysical, structural NMR, and molecular docking characterization.

I

Intervention

D-peptide inhibitor ER-DRI titrated at molar ratios of 1:2, 1:4, and 1:8 against 100 μmol/L 15N-labeled EV-A71 3A(1-57) protein, evaluated via 1H-15N HSQC NMR spectroscopy and molecular docking.

C

Comparator

Apo-form 15N-labeled EV-A71 3A(1-57) protein in the absence of ER-DRI (1:0 molar ratio).

O

Outcome

Backbone resonance assignments, secondary structure prediction, 15N relaxation dynamics (R1, R2, hNOE), rotational correlation time (τc), and chemical shift line broadening upon ER-DRI titration.surrogate

Elucidates the structural dynamics of apo-form EV-A71 3A protein and demonstrates that the peptide inhibitor ER-DRI competitively binds its conserved homodimerization interface, aiding the rational design of broad-spectrum anti-enteroviral therapeutics.

Limitations

75% (42/56) of backbone amide 1H-15N pairs were assigned, with overlapped peaks or low signal-to-noise ratios excluded from relaxation analysis
Further experimental investigation on the binding details is needed to elucidate the exact target site of 3A(1-57) on ER-DRI
Unassigned residues and residues with overlapped peaks or low signal-to-noise ratios (only 37 non-proline residues analyzed for 15N relaxation)
Molecular docking showed the helix motif of 3A(1-57) bound alternatively to the TAT peptide segment of ER-DRI rather than the anticipated helix-mimicking part, requiring further experimental investigation

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Implication

ER-DRI binding supports targeted EV-A71 inhibition; leaves open in vivo efficacy and clinical translation.

Abstract

• Our results disclosed a dihelical structure of Enterovirus A71 3A 1–57 protein in apo form. • We depicted rigid helices and a unique flexible C-terminus for apo-form 3A 1–57 . • This study revealed a competitive binding-based molecular mechanism underlying inhibition of dimeric 3A by ER-DRI.

Discussion

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Cite This Study

Liu et al. (2023) studied Enterovirus A71 (EV-A71) infection / Hand, foot, and mouth disease (HFMD). D-peptide inhibitor ER-DRI vs. Untreated apo-form 15N-labeled EV-A71 3A(1-57) protein (1:0 molar ratio) was evaluated on Structural dynamics and intermolecular binding interactions between EV-A71 3A(1-57) protein and ER-DRI via NMR spectroscopy and molecular docking. D-peptide ER-DRI competitively binds to the dimeric α-helix interface of EV-A71 3A(1-57) protein (apparent dimer Mw 16.78 kDa, τc 10.49 ns) to inhibit homodimerization.

synapsesocial.com/papers/6a7f355ad9ea295be639d63d https://doi.org/10.1016/j.virs.2023.09.004

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Structural insight into EV-A71 3A protein and its interaction with a peptide inhibitor

Figures

Key Points

Structured PICO

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

Structural insight into EV-A71 3A protein and its interaction with a peptide inhibitor

Figures

Key Points

Structured PICO

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

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