6S6E

Crystal structure of the engineered ancestor of haloalkane dehalogenases and Renilla luciferase (AncHLD-RLuc I161_F162PinsL)

6S6E の概要

• エントリーDOI: 10.2210/pdb6s6e/pdb
• 分子名称: Engineered ancestor of haloalkane dehalogenases and Renilla luciferase (AncHLD-RLuc I161_F162PinsL) (2 entities in total)
• 機能のキーワード: engineered ancestral enzyme, bioluminscence, coelenterazine-utilizing enzyme, luminescent protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 68411.62

構造登録者

Schenkmayerova, A.,Damborsky, J.,Marek, M. (登録日: 2019-07-03, 公開日: 2021-01-27, 最終更新日: 2024-01-24)

主引用文献

Schenkmayerova, A.,Pinto, G.P.,Toul, M.,Marek, M.,Hernychova, L.,Planas-Iglesias, J.,Daniel Liskova, V.,Pluskal, D.,Vasina, M.,Emond, S.,Dorr, M.,Chaloupkova, R.,Bednar, D.,Prokop, Z.,Hollfelder, F.,Bornscheuer, U.T.,Damborsky, J.
Engineering the protein dynamics of an ancestral luciferase.
Nat Commun, 12:3616-3616, 2021

PubMed Abstract: 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 which catalyses both dehalogenase and luciferase reactions. Insertion-deletion (InDel) backbone mutagenesis of Anc 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.

PubMed: 34127663
DOI: 10.1038/s41467-021-23450-z

実験手法

X-RAY DIFFRACTION (2.001 Å)