Protein dynamics at slow timescales in engineered ß-lactamases does not limit evolvability
September 15-19, 2019
Understanding the underlying mechanisms in the evolution of new protein functions is key to better directing enzyme engineering efforts. Intragenic epistasis (the non-additive interaction of mutations affecting function) is a key feature of protein evolution. For example, in TEM-1 ß-lactamase, the mutations E104K and G238S show positive epistasis in that their combination show a greater than expected increase in antibiotic resistance1. Here, we aim to understand the impact of protein dynamics at slow timescales on epistasis. Large conformational rearrangements associated with ligand-binding, turnover of substrate or allostery occur at this timescale2. The readily evolvable antibiotic-resistant TEM-1 ß-lactamase and two of its variants that are more dynamic at slow timescales serve as models for this study. Our models show similar catalytic activity and substrate recognition, thermal stability, as well as conserved motions in fast (ps-ns) and intermediate (ns-ms) timescales but different motions at slow timescales (ms-ms)123.
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Lorea Alejaldre, Ferran Sancho, Claudèle Lemay-St-Denis, Joelle N. Pelletier, Adem H-Parisien, and Victor Guallar, "Protein dynamics at slow timescales in engineered ß-lactamases does not limit evolvability" in "Enzyme Engineering XXV", Huimin Zhao, University of Illinois at Urbana-Champaign, USA John Wong, Pfizer, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/enzyme_xxv/50