Structural and functional investigation of mycobacterial HflX protein and its mutational hotspots annotation by in silico approach

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) remains a critical burden on global health. GTP binding proteins play a considerable role in the cell signaling processes of Mtb. The present manuscript elaborates important aspects of a mycobacterial gene Rv2725c (HfIX), a probable GTP binding protein. The HflX protein contains four GTP binding motifs, which postulates its role in GTP binding activity. In silico assessment of the HflX protein was done by various computational tools. A homology modelling perspective has been employed to define the tertiary structure of the protein using the SWISS-MODEL server and further validated by the RAMPAGE server. In addition, secondary structure, protein-protein and protein-chemical interactions, phosphorylation site prediction, ligand binding sites, molecular docking and mutational investigations were performed to give more recognition to HflX. Mycobacterial HflX protein is predicted to be present in the cytoplasmic compartment, interacts with the ribosome in a GTP-dependent manner, and helps in translational processes. Hence, the present study provides a way to understand its normal functioning and basic biology, which can enhance the information for developing novel therapeutic targeting approaches for HflX protein.