# Maintenance of native-like protein dynamics may not be required for engineering functional proteins

Sophie M.C. Gobeil, Christopher M. Clouthier, Jaeok Park, Donald Gagné,

Albert M. Berghuis, Nicolas Doucet, Joelle N. Pelletier

► To cite this version:

Sophie M.C. Gobeil, Christopher M. Clouthier, Jaeok Park, Donald Gagné, Albert M. Berghuis, et al. Maintenance of native-like protein dynamics may not be required for engineering functional proteins. Chemistry and Biology, 2014, 21 (10), pp.1330-1340. 10.1016/j.chembiol.2014.07.016. hal-01196413

HAL Id: hal-01196413

https://hal.science/hal-01196413

Submitted on 14 Sep 2015

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# Maintenance of native-like proteindynamics may not be required for engineering functional proteins.

Gobeil SM¹, Clouthier CM², Park J³, Gagné D⁴, Berghuis AM³, Doucet N⁴, Pelletier JN⁵.

## Author information

¹PROTEO Network, Université Laval, Québec QC G1V 0A6, Canada; Département de Biochimie, Université de Montréal, Montréal QC H3T 1J4, Canada.

²PROTEO Network, Université Laval, Québec QC G1V 0A6, Canada; Département de Chimie, Université de Montréal, Montréal QC H3T 1J4, Canada.

³PROTEO Network, Université Laval, Québec QC G1V 0A6, Canada; Department of Biochemistry and Department of Microbiology and Immunology, McGill University, Montreal QC H3G 1Y6, Canada; GRASP Network, McGill University, Montréal QC H3G 1Y6, Canada.

⁴PROTEO Network, Université Laval, Québec QC G1V 0A6, Canada; GRASP Network, McGill University, Montréal QC H3G 1Y6, Canada; INRS-Institut Armand-Frappier, Université du Québec, Laval QC H7V 1B7, Canada.

⁵PROTEO Network, Université Laval, Québec QC G1V 0A6, Canada; Département de Biochimie, Université de Montréal, Montréal QC H3T 1J4, Canada; Département de Chimie, Université de Montréal, Montréal QC H3T 1J4, Canada; Center for Green Chemistry and Catalysis (CCVC), Montréal QC H3A 0B8, Canada. Electronic address: joelle.pelletier@umontreal.ca.

## Abstract

Proteins are dynamicsystems, and understanding dynamics is critical for fully understanding protein function. Therefore, the question of whether laboratory engineering has an impact on prote dynamics is of general interest. Here, we demonstrate that two homologous, naturally evolved enzymes with high degrees of structural and functional conservation also exhibit conserved dynamics. Their similar set of slow timescale dynamics is highly restricted, consistent with the evolutionary conservation of a functionally important feature. However, we also show that dynamics of a laboratory-engineered chimeric enzyme obtained by recombination of the two homologous exhibits striking difference on the millisecond timescale, despite function and high-resolution crystal structure (1.05 Å) being conserved. The laboratory-engineered chimera is thus functionally tolerant to modified dynamics on the timescale of catalytic turnover. Tolerance to dynamic variation implies that maintenance of native-like prote dynamics may not be required when engineering functional proteins.