CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2025 , Vol. 43 >Issue 2: 192 - 197

DOI: https://doi.org/10.12140/j.issn.1000-7423.2025.02.007

ORIGINAL ARTICLES

Analysis of the key sites of Plasmodium vivax VIR14 protein interacting with ICAM-1 receptor

  • YANG Wanxuan ,
  • SHEN Haimo ,
  • CHEN Shenbo ,
  • CHEN Junhu
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  • 1 School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou 310013, Zhejiang, China
    2 National Institute of Parasitic Diseases, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; Chinese Center for Diseases Control and Prevention (Chinese Center for Tropical Diseases Research); NHC Key Laboratory of Parasite and Vector Biology; WHO Collaborating Centre for Tropical Diseases; National Centre for International Research on Tropical Diseases, Shanghai 200025, China

Received date: 2025-01-27

  Revised date: 2025-03-27

  Online published: 2025-04-27

Supported by

Project of Shanghai Science and Technology Commission(18490741100)

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Abstract

Objective To investigate the binding of Plasmodium vivax VIR14 protein to intercellular cell adhesion molecule-1 (ICAM-1) receptor and identify key binding sites. Methods The recombinant plasmid pET28a-VIR14 was transformed into Escherichia coli BL21 (DE3). The expression levels and purity of VIR14 protein were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Binding of VIR14 protein or peptides to ICAM-1 was verified using surface plasmon resonance (SPR). Structures of VIR14 and ICAM-1 (28-480 aa) were predicted, and complex modeling scores (pTM and pLDDT) were integrated. Docking analysis was performed using ClusPro 2.0 to evaluate the interactions between top-scoring VIR14 and ICAM-1 models. Docking sites were analyzed using PDBePISA and Pymol 3.0. Amino acid polymorphisms at binding sites were examined using global isolate data. Key interaction sites were analyzed via epitope competition inhibition assays. Results SDS-PAGE confirmed successful expression and purification of VIR14 protein, with a relactive molecular mass (Mr) of 58 000. SPR showed concentration-dependent binding of VIR14 to ICAM-1, with an affinity of 15.16 μmol/L. According to structural predictions from Alphafold2, the optimal model of the VIR14 protein had a pLDDT score of 78.2 and a pTM score of 0.73. In contrast, the optimal model of the ICAM-1 receptor showed a pLDDT score of 93.8 and a pTM score of 0.65. The regions where polar bonds formed between the VIR14 and ICAM-1 proteins were predominantly located in the C-terminal region of the protein, specifically between the 212 and 406 amino acids. Among the two peptides containing the predicted binding sites, only peptide P2 bound to ICAM-1, with an affinity of 48.99 μmol/L. Competitive inhibition experiments indicated that P2 could inhibit the binding of VIR14 protein to ICAM-1. After replacing the side chains of the four key residues (ASP-310, ARG-314, LYS-316, and ASP-318) with alanine, P2 lost its inhibitory function. P2 was highly conserved across 338 P. vivax global isolates. Conclusion The VIR14 protein of P. vivax exhibits tight binding to the ICAM-1 receptor, with four amino acid residues playing essential roles in this interaction. These key sites demonstrate high conservation across P. vivax global isolates.

Key words： Plasmodium vivax; VIR protein; Surface plasmon resonance; Alphafold2; Genetic diversity

Cite this article

YANG Wanxuan , SHEN Haimo , CHEN Shenbo , CHEN Junhu . Analysis of the key sites of Plasmodium vivax VIR14 protein interacting with ICAM-1 receptor[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025 , 43(2) : 192 -197 . DOI: 10.12140/j.issn.1000-7423.2025.02.007

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