title: Inhibitory effects of a peptide-fusion protein (Latarcin-PAP1-Thanatin) against chikungunya virus.
abstract: Chikungunya virus (CHIKV) outbreaks have led to a serious economic burden, as the available treatment strategies can only alleviate disease symptoms, and no effective therapeutics or vaccines are currently available for human use. Here, we report the use of a new cost-effective approach involving production of a recombinant antiviral peptide-fusion protein that is scalable for the treatment of CHIKV infection. A peptide-fusion recombinant protein LATA-PAP1-THAN that was generated by joining Latarcin (LATA) peptide with the N-terminus of the PAP1 antiviral protein, and the Thanatin (THAN) peptide to the C-terminus, was produced in Escherichia coli as inclusion bodies. The antiviral LATA-PAP1-THAN protein showed 89.0% reduction of viral plaque formation compared with PAP1 (46.0%), LATA (67.0%) or THAN (79.3%) peptides alone. The LATA-PAP1-THAN protein reduced the viral RNA load that was 0.89-fold compared with the untreated control cells. We also showed that PAP1 resulted in 0.44-fold reduction, and THAN and LATA resulting in 0.78-fold and 0.73-fold reductions, respectively. The LATA-PAP1-THAN protein inhibited CHIKV replication in the Vero cells at an EC50 of 11.2μg/ml, which is approximately half of the EC50 of PAP1 (23.7μg/ml) and protected the CHIKV-infected mice at the dose of 0.75mg/ml. We concluded that production of antiviral peptide-fusion protein in E. coli as inclusion bodies could accentuate antiviral activities, enhance cellular internalisation, and could reduce product toxicity to host cells and is scalable to epidemic response quantities.
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: rothan@um.edu.my.
authors.forename: Hussin A
authors.initials: HA
authors.lastname: Rothan
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: hirbodb83@gmail.com.
authors.forename: Hirbod
authors.initials: H
authors.lastname: Bahrani
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: shankarem@um.edu.my.
authors.forename: Esaki M
authors.initials: EM
authors.lastname: Shankar
authors.affiliation: Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: noorsaadah@um.edu.my.
authors.forename: Noorsaadah Abd
authors.initials: NA
authors.lastname: Rahman
authors.affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: rohana@um.edu.my.
authors.forename: Rohana
authors.initials: R
authors.lastname: Yusof
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D000998:Antiviral Agents / Q000494:pharmacology / Q000627:therapeutic use*; D065632:Chikungunya Fever / Q000188:drug therapy / Q000517:prevention & control*; D002646:Chikungunya virus / Q000187:drug effects* / Q000502:physiology; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008813:Mice, Inbred ICR; D008826:Microbial Sensitivity Tests; D000073718:Pancreatitis-Associated Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D054789:Ribosome Inactivating Proteins, Type 1 / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D013111:Spider Venoms / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D016896:Treatment Outcome; D014709:Vero Cells; D019562:Viral Load; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects
chemical_list: D023181:Antimicrobial Cationic Peptides; D000998:Antiviral Agents; D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D013111:Spider Venoms; C531159:latarcin 2a, Lachesana tarabaevi; C098076:thanatin; C024275:pokeweed antiviral protein
keywords: Antiviral activity; Chikungunya virus; Fusion protein; Latarcin; Tachyplesin1; Thanatin