Abstract
Previously, a hypothetical protein (HP) termed Bleg1_2437 (currently named Bleg1_2478) from Bacillus lehensis G1 was discovered to be an evolutionary divergent B3 subclass metallo-β-lactamase (MBL). Due to the scarcity of clinical inhibitors for B3 MBLs and the divergent nature of Bleg1_2478, this study aimed to design and characterise peptides as inhibitors against Bleg1_2478. Through in silico docking, RSWPWH and SSWWDR peptides with comparable binding energy to ampicillin were obtained. In vitro assay results showed RSWPWH and SSWWDR inhibited the activity of Bleg1_2478 by 50% at concentrations as low as 0.90 µM and 0.50 µM, respectively. At 10 µM of RSWPWH and 20 µM of SSWWDR, the activity of Bleg1_2478 was almost completely inhibited. Isothermal titration calorimetry (ITC) analyses showed slightly improved binding properties of the peptides compared to ampicillin. Docked peptide–protein complexes revealed that RSWPWH bound near the vicinity of the Bleg1_2478 active site while SSWWDR bound at the center of the active site itself. We postulate that the peptides caused the inhibition of Bleg1_2478 by reducing or blocking the accessibility of its active site from ampicillin, thus hampering its catalytic function.
Highlights
ObjectivesDue to the scarcity of clinical inhibitors for B3 MBLs and the divergent nature of Bleg1_2478, this study aimed to design and characterise peptides as inhibitors against
In view of limited inhibitors against B3 MBLs to date and the evolutionary divergent nature of Bleg1_2478, this study aims to design inhibitory peptides against Bleg1_2478 B3 subclass MBL and characterise their inhibitory potential and properties through in vitro and in silico approaches
Docking results suggested that there was only one specific binding site in Bleg1_2478 for ampicillin, which was at the active site which houses the metal-binding residues important for its catalytic activity (Figure 1)
Summary
Due to the scarcity of clinical inhibitors for B3 MBLs and the divergent nature of Bleg1_2478, this study aimed to design and characterise peptides as inhibitors against. In view of limited inhibitors against B3 MBLs to date and the evolutionary divergent nature of Bleg1_2478, this study aims to design inhibitory peptides against Bleg1_2478 B3 subclass MBL and characterise their inhibitory potential and properties through in vitro and in silico approaches
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