Fig 1.
Proteome exploration of Vibrio parahaemolyticus to identify novel drug targets.
Fig 2.
Flow chart summarizing the protocols over multi-epitope subunit vaccine development against V. parahaemolyticus through reverse vaccinology approach.
Table 1.
Subtractive proteomics analysis scheme.
Table 2.
Pathogen specific essential cytoplasmic proteins as novel therapeutic targets.
Fig 3.
Investigation of PPIs through STRING v10.5 server; (A) UDP-N-acetylmuramoyl-L-alanyl-D-glutamate—2,6-diaminopimelate ligase (murE), (B) Trigger factor (tig).
Table 3.
Predicted therapeutic targets (novel cytoplasmic proteins) showing virulent properties.
Table 4.
Top 10 metabolites predicted as suitable drug candidates against VIBPA Type II secretion system protein L and VIBPA Putative fimbrial protein Z.
Table 5.
ADME profiling of top drug candidates.
Table 6.
Novel vaccine targets proteins showing higher antigenicity.
Fig 4.
Population coverage analysis of (A) VIPBA putative sensor histidine protein kinase UhpB, and (B) Flagellar hook-associated protein.
Table 7.
Predicted final CTL and HTL epitopes of histidine protein kinase and flagellar hook-associated protein.
Table 8.
Binding energy of predicted epitopes with selected MHC class I and MHC class II molecules generated from molecular docking by AutoDock.
Fig 5.
Solubility prediction of vaccine constructs.
(A) Solubility prediction of designed vaccine construct V1 using via Protein-sol server, and (B) prediction of polar, nonpolar, hydrophobic and aromatic regions.
Fig 6.
Tertiary structure prediction and validation of vaccine protein V1.
(A) Tertiary structure of modeled construct V1, (B) Ramachandran plot analysis of vaccine protein V1.
Fig 7.
Docked complex of vaccine construct V1 with human TLR 1/2 heterodimer.
(A) Cartoon format, and (B) Ball structure.
Fig 8.
Molecular dynamics simulation of vaccine protein V1-TLR8 complex.
Stability of the protein-protein complex was investigated through (A) mobility, (B) B-factor, (C) deformability, (D) eigenvalue, (E) covariance and (F) elastic network analysis.
Fig 9.
In silico restriction cloning of the gene sequence of construct V1 into pET28a(+) expression vector; (A) Restriction digestion of the vector pET28a(+) and construct V1 with BglI and ApaI (B) Inserted desired fragment (V1 Construct) between ApaI (1334) and BglI (2452) indicated in red color.