Computational Design of Phosphotriesterase Improves V‐Agent Degradation Efficiency

• Published in: ChemistryOpen (Weinheim) Vol. 13; no. 7; pp. e202300263 - n/a
• Main Authors: Kronenberg, Jacob, Chu, Stanley, Olsen, Andrew, Britton, Dustin, Halvorsen, Leif, Guo, Shengbo, Lakshmi, Ashwitha, Chen, Jason, Kulapurathazhe, Maria Jinu, Baker, Cetara A., Wadsworth, Benjamin C., Van Acker, Cynthia J., Lehman, John G., Otto, Tamara C., Renfrew, P. Douglas, Bonneau, Richard, Montclare, Jin Kim
• Format: Journal Article
• Language: English
• Published: Germany John Wiley & Sons, Inc 01.07.2024; John Wiley and Sons Inc; Wiley-VCH
• Subjects: Catalysis; Cholinesterase Inhibitors - chemistry; Cholinesterase Inhibitors - metabolism; computational design; Efficiency; enzyme engineering; Enzymes; Hydrolysis; Models, Molecular; Mutation; nanoscavengers; nerve agents; Organophosphates; Organothiophosphorus Compounds - chemistry; Organothiophosphorus Compounds - metabolism; Phosphoric Triester Hydrolases - chemistry; Phosphoric Triester Hydrolases - metabolism; Scavengers; Spectrum analysis; enzyme engineering; computational design; nerve agents; organophosphates; nanoscavengers
• ISSN: 2191-1363; 2191-1363
• DOI: 10.1002/open.202300263

Organophosphates (OPs) are a class of neurotoxic acetylcholinesterase inhibitors including widely used pesticides as well as nerve agents such as VX and VR. Current treatment of these toxins relies on reactivating acetylcholinesterase, which remains ineffective. Enzymatic scavengers are of interest for their ability to degrade OPs systemically before they reach their target. Here we describe a library of computationally designed variants of phosphotriesterase (PTE), an enzyme that is known to break down OPs. The mutations G208D, F104A, K77A, A80V, H254G, and I274N broadly improve catalytic efficiency of VX and VR hydrolysis without impacting the structure of the enzyme. The mutation I106 A improves catalysis of VR and L271E abolishes activity, likely due to disruptions of PTE's structure. This study elucidates the importance of these residues and contributes to the design of enzymatic OP scavengers with improved efficiency. Organophosphates (OPs) are potent neurotoxins whose current remedies are not very effective. Here we design and characterize variants of the enzyme phosphotriesterase, which can degrade OPs. We report mutations improving catalytic efficiency between 2‐ and 5‐fold and confirm folding and stability of the resulting variants. These findings are a step towards improved OP bioscavengers.