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@article{48251,
   author = {Schenkmayerová, Andrea and Rangel Pamplona Pizarro Pinto, José Gaspar and Toul, Martin and Marek, Martin and Hernychová, Lenka and Planas Iglesias, Joan and Lišková, Veronika and Pluskal, Daniel and Vašina, Michal and Emond, Stephane and Dörr, Mark and Chaloupková, Radka and Bednář, David and Prokop, Zbyněk and Hollfelder, Florian and Bornscheuer, Uwe T. and Damborský, Jiří},
   article_location = {London},
   article_number = {1},
   doi = {http://dx.doi.org/10.1038/s41467-021-23450-z},
   keywords = {Enzymes; Hydrolases; Protein design; X-ray crystallography},
   language = {eng},
   issn = {2041-1723},
   journal = {Nature Communications},
   title = {Engineering the protein dynamics of an ancestral luciferase},
   url = {https://doi.org/10.1038/s41467-021-23450-z},
   volume = {12},
   year = {2021}
}

TY  - JOUR
ID  - 48251
AU  - Schenkmayerová, Andrea - Rangel Pamplona Pizarro Pinto, José Gaspar - Toul, Martin - Marek, Martin - Hernychová, Lenka - Planas Iglesias, Joan - Lišková, Veronika - Pluskal, Daniel - Vašina, Michal - Emond, Stephane - Dörr, Mark - Chaloupková, Radka - Bednář, David - Prokop, Zbyněk - Hollfelder, Florian - Bornscheuer, Uwe T. - Damborský, Jiří
PY  - 2021
TI  - Engineering the protein dynamics of an ancestral luciferase
JF  - Nature Communications
VL  - 12
IS  - 1
SP  - 3616
EP  - 3616
PB  - Nature Publishing Group
SN  - 2041-1723
KW  - Enzymes
KW  - Hydrolases
KW  - Protein design
KW  - X-ray crystallography
UR  - https://doi.org/10.1038/s41467-021-23450-z
N2  - Directed evolution commonly relies on point mutations but InDels frequently occur in evolution. Here the authors report a protein-engineering framework based on InDel mutagenesis and fragment transplantation resulting in greater catalysis and longer glow-type bioluminescence of the ancestral luciferase. Protein dynamics are often invoked in explanations of enzyme catalysis, but their design has proven elusive. Here we track the role of dynamics in evolution, starting from the evolvable and thermostable ancestral protein Anc(HLD-RLuc) which catalyses both dehalogenase and luciferase reactions. Insertion-deletion (InDel) backbone mutagenesis of Anc(HLD-RLuc) challenged the scaffold dynamics. Screening for both activities reveals InDel mutations localized in three distinct regions that lead to altered protein dynamics (based on crystallographic B-factors, hydrogen exchange, and molecular dynamics simulations). An anisotropic network model highlights the importance of the conformational flexibility of a loop-helix fragment of Renilla luciferases for ligand binding. Transplantation of this dynamic fragment leads to lower product inhibition and highly stable glow-type bioluminescence. The success of our approach suggests that a strategy comprising (i) constructing a stable and evolvable template, (ii) mapping functional regions by backbone mutagenesis, and (iii) transplantation of dynamic features, can lead to functionally innovative proteins.
ER  - 

SCHENKMAYEROVÁ, Andrea, José Gaspar RANGEL PAMPLONA PIZARRO PINTO, Martin TOUL, Martin MAREK, Lenka HERNYCHOVÁ, Joan PLANAS IGLESIAS, Veronika LIŠKOVÁ, Daniel PLUSKAL, Michal VAŠINA, Stephane EMOND, Mark DÖRR, Radka CHALOUPKOVÁ, David BEDNÁŘ, Zbyněk PROKOP, Florian HOLLFELDER, Uwe T. BORNSCHEUER a Jiří DAMBORSKÝ. Engineering the protein dynamics of an ancestral luciferase. \textit{Nature Communications}. London: Nature Publishing Group, 2021, roč.~12, č.~1, s.~3616-3631. ISSN~2041-1723. Dostupné z: https://dx.doi.org/10.1038/s41467-021-23450-z.