Characterization of Helicobacter pylori immunoreactive proteins NusB, isoprenyl transferase, and hypothetical protein via immunoproteomics and molecular modeling approaches

The microaerophilic Gram-negative bacterium H. pylori is associated with various gastric complications and affects nearly half of the global population. Current sero-diagnostic methods for H. pylori diagnosis are often insensitive or lack specificity. This study aimed to detect H. pylori immunoreactive proteins to improve diagnostic tools. H. pylori isolates from biopsy samples were characterized using biochemical and molecular techniques. An immunoproteomics approach involving immunoprecipitation and mass spectrometry identified three immunoreactive proteins: Transcription antitermination protein NusB, Isoprenyl transferase, and a hypothetical protein associated with a transposase gene. Bioinformatics analysis revealed that these proteins are involved in RNA binding, termination of DNA-templated transcription, cell and energy metabolism, transferase activity, regulation, and ribosomal biosynthesis pathways. CD4 T cell and Class-I immunogenicity predictions highlighted NusB's strong potential to stimulate an immune response. Immune simulations demonstrated robust antibody production, particularly in response to NusB. Additionally, molecular docking studies with phenolic compounds (Gnetol, Isohomovanillic acid, Licoisoflavone A, and Chrysosplenol D) against the three proteins, followed by molecular dynamics (MD) simulations, confirmed the stability and favorable interactions of these protein-phenolic compound complexes. This integrative approach, combining immunoproteomics, bioinformatics, molecular docking, and MD simulations, underscores the potential of these immunoreactive proteins for vaccine development and improved diagnostic methods.