Identification, Characterization and Phylogenetic Analysis of a Novel Salivary Apyrase Gene from the Arbovirus Vector Culex antennatus

Novel Salivary Apyrase Gene from the Arbovirus Vector Culex antennatus

Authors

  • AlaaEddeen Seufi Cairo University
  • Tanani Zoology Department, Faculty of Science, Al-Azhar University, Egypt.
  • Alaa G. Al-Daly Zoology Department, Faculty of Science, Al-Azhar University, Egypt.
  • Ramy E. M. El-Ansary Zoology Department, Faculty of Science, Al-Azhar University, Egypt.
  • Fatma H. Galal Biology Department, College of Science, Jouf University, Aljouf, Sakaka, Saudi Arabia; Department of Entomology, Faculty of Science , Cairo University, Giza, Egypt.

Keywords:

Culex antennatus, Salivary Apyrase, Hematophagy, Ca2 -Dependent NTPDase, Molecular Docking, Vector-Host Interactions

Abstract

Hematophagous arthropods rely on complex bioactive salivary secretions to counteract host physiological defenses during blood-feeding. Among these salivary proteins, ecto-apyrases (nucleoside triphosphate diphosphohydrolases/ 5'-nucleotidases) play a vital role by hydrolyzing extracellular adenosine triphosphate (ATP) and adenosine diphosphate (ADP) released from damaged vascular tissues, thereby preventing platelet aggregation, neutrophil recruitment, and pain signaling. In this study, we identified, cloned, and structurally characterized a novel salivary apyrase cDNA (CxanApy) from the primary arbovirus vector Culex antennatus. Differential display RT-PCR (DD-PCR) transcriptomic profiling across varied developmental, nutritional, and post-ingestion phases demonstrated inducible upregulation during active probing and early blood-meal processing. Using 5'-RACE and gene-specific PCR, a 636 bp cDNA encoding a 522 bp open reading frame (ORF) was isolated. The deduced 173-amino acid precursor includes an N-terminal signal peptide (M1- A16) whose cleavage yields a mature, soluble ~17.3 kDa secreted enzyme with an acidic pI of 5.12. In silico topological mapping, homology modeling, and docking simulations revealed a classic α/β-sandwich fold containing five highly conserved Apyrase Conserved Regions (ACRs 1–5), an intramolecular disulfide loop (Cys48- Cys53), and active-site catalytic triads (His36- Asp33, His77- Asp80, His112- Asp120) capable of coordinating Ca2+/ Mn2+ divalent cations for nucleoside hydrolysis. Phylogenetic reconstruction positioned CxanApy within a monophyletic Culex lineage closely aligned with Culex quinquefasciatus and Culex pipiens orthologs. These insights into vector salivary biochemistry highlight potential molecular targets for disrupting blood-feeding efficiency and arboviral transmission cascades.

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Published

2024-09-13

How to Cite

Seufi, A., Tanani, M. A., Al-Daly , A. G., El-Ansary, R. E. M. ., & Galal, F. H. (2024). Identification, Characterization and Phylogenetic Analysis of a Novel Salivary Apyrase Gene from the Arbovirus Vector Culex antennatus: Novel Salivary Apyrase Gene from the Arbovirus Vector Culex antennatus. WAS Science Nature (WASSN) ISSN: 2766-7715, 7(1), 1–22. Retrieved from https://www.worldascience.com/journals/index.php/wassn/article/view/54

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