Abstract: Background and Objective: A comprehensive study of Pseudomonas aeruginosa proteomics found that 25% of proteins are retained as hypothetical proteins whose functions have not yet been ultimately determined. In this study, it’s attempted to assign a particular role to one such hypothetical protein PA2373 for which no experimental knowledge is currently available. Materials and Methods: To achieve this, the newest versions of bioinformatics tools was applied that provide various information, such as regarding amino acid sequences, protein families, sequence-function relationships and motifs. To identify homologous proteins, sequence similarities were searched using accessible bioinformatics databases. Results: Obtained results showed that PA2373 has two functional domains, including VI_Rhs_Vgr and 5 superfamilies. Functional annotation showed that PA2373 could be a Vgr protein and type VI secretion system. In addition, protein-protein interactions of selected hypothetical proteins show that certain functional partners play a significant role in pathogen survival. Conclusion: This study's findings may help better understand the mechanisms of virulence, drug resistance and pathogenesis of P. aeruginosa infections and to innovate the treatment strategies subsequently.