Supercharged Phosphotriesterase for improved Paraoxon activity.

Autor:	Kronenberg J; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Britton D; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Halvorsen L; Center for Genomics and Systems Biology, New York University, New York, New York 10003, USA., Chu S; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Kulapurathazhe MJ; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Chen J; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Lakshmi A; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA., Renfrew PD; Center for Genomics and Systems Biology, New York University, New York, New York 10003, USA., Bonneau R; Center for Genomics and Systems Biology, New York University, New York, New York 10003, USA.; Center for Computational Biology, Flatiron Institute, Simons Foundation, New York, New York 10010, USA.; Courant Institute of Mathematical Sciences, Computer Science Department, New York University, New York, New York 10009, USA., Montclare JK; Department of Chemical and Biomolecular Engineering, New York University Tandon School of Engineering, Brooklyn, New York 11201, USA.; Bernard and Irene Schwartz Center for Biomedical Imaging, Department of Radiology, New York University School of Medicine, New York, New York 10016, USA.; Department of Chemistry, New York University, New York, New York 10012, USA.; Department of Biomaterials, New York University College of Dentistry, New York, New York 10010, USA.; Department of Biomedical Engineering, New York University, New York, NY 11201, USA.
Jazyk:	angličtina
Zdroj:	Protein engineering, design & selection : PEDS [Protein Eng Des Sel] 2024 Jan 29; Vol. 37.
DOI:	10.1093/protein/gzae015
Abstrakt:	Phosphotriesterases (PTEs) represent a class of enzymes capable of efficient neutralization of organophosphates (OPs), a dangerous class of neurotoxic chemicals. PTEs suffer from low catalytic activity, particularly at higher temperatures, due to low thermostability and low solubility. Supercharging, a protein engineering approach via selective mutation of surface residues to charged residues, has been successfully employed to generate proteins with increased solubility and thermostability by promoting charge-charge repulsion between proteins. We set out to overcome the challenges in improving PTE activity against OPs by employing a computational protein supercharging algorithm in Rosetta. Here, we discover two supercharged PTE variants, one negatively supercharged (with -14 net charge) and one positively supercharged (with +12 net charge) and characterize them for their thermodynamic stability and catalytic activity. We find that positively supercharged PTE possesses slight but significant losses in thermostability, which correlates to losses in catalytic efficiency at all temperatures, whereas negatively supercharged PTE possesses increased catalytic activity across 25°C-55°C while offering similar thermostability characteristic to the parent PTE. The impact of supercharging on catalytic efficiency will inform the design of shelf-stable PTE and criteria for enzyme engineering. (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)
Databáze:	MEDLINE
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