Identification of HPr kinase/phosphorylase inhibitors: novel antimicrobials against resistant Enterococcus faecalis

Abstract

Enterococcus faecalis, a gram-positive bacterium, is among the most common nosocomial pathogens due to its limited susceptibility to antibiotics and its reservoir of the genes coding for virulence factors. Bacterial enzymes such as kinases and phosphorylases play important roles in diverse functions of a bacterial cell and, thus, are potential antibacterial drug targets. In Gram-positive bacteria, HPr Kinase/Phosphorylase (HPrK/P), a bifunctional enzyme is involved in the regulation of carbon catabolite repression by phosphorylating/dephosphorylating the histidine-containing phosphocarrier protein (HPr) at Ser46 residue. Deficiencies in HPrK/P function leads to severe defects in bacterial growth. This study aimed at identifying novel inhibitors of E. faecalis HPrK/P from a commercial compound library using structure-based virtual screening. The hit molecules were purchased and their effect on enzyme activity and growth of resistant E. faecalis was evaluated in vitro. Furthermore, docking and molecular dynamics simulations were performed to study the interactions of the hit compounds with HPrK/P. Among the identified hit molecules, two compounds inhibited the phosphorylation of HPr as well as significantly reduced the growth of resistant E. faecalis in vitro. These identified potential HPrK/P inhibitors open new research avenues towards the development of novel antimicrobials against resistant Gram-positive bacteria.

Proteomics and metabolomics analyses of Streptococcus agalactiae isolates from human and animal sources

“…Kinases are crucial for bacterial growth, biofilm formation, and antibiotic resistance. Recent studies have reported the important role of penicillin-binding protein and serine/threonine kinase-associated (PASTA) kinases in regulating resistance to β-lactam antibiotics 40 , 41 . Hence, bacterial kinases have received considerable attention as prospective targets for the discovery of novel antimicrobial agents 40 .…”

“…Recent studies have reported the important role of penicillin-binding protein and serine/threonine kinase-associated (PASTA) kinases in regulating resistance to β-lactam antibiotics 40 , 41 . Hence, bacterial kinases have received considerable attention as prospective targets for the discovery of novel antimicrobial agents 40 . Interestingly, animal strains uniquely contained histidine kinase (cssS, maeK, dcuS_2, and dcuS) and HPr kinase/phosphorylase (HPrK/P) proteins (isolates code no.…”