Automated Engineering Protein Dynamics via Loop Grafting: Improving Renilla Luciferase Catalysis

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F25%3A00082465" target="_blank" >RIV/00159816:_____/25:00082465 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/25:00140815

Result on the web

<a href="https://pubs.acs.org/doi/10.1021/acscatal.4c06207" target="_blank" >https://pubs.acs.org/doi/10.1021/acscatal.4c06207</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acscatal.4c06207" target="_blank" >10.1021/acscatal.4c06207</a>

Result language

angličtina

Original language name

Automated Engineering Protein Dynamics via Loop Grafting: Improving Renilla Luciferase Catalysis

Original language description

Engineering protein dynamics is a challenging and unsolved problem in protein design. Loop transplantation or loop grafting has been previously employed to transfer dynamic properties between proteins. We recently released a LoopGrafter Web server to execute the loop grafting task, employing eight computational tools and one database. The LoopGrafter method relies on the prediction of the local dynamic behavior of the elements to be transplanted and has successfully reconstructed previously engineered sequences. However, it was unclear whether catalytically competitive previously uncharacterized designs could be obtained by this method. Here, we address this question, showing how LoopGrafter generates viable loop-grafted chimeras of luciferases, how these chimeras encompass the activity of interest and unique kinetic properties, and how all this process is done fully automatically and agnostic of any previous knowledge. All constructed designs proved to be catalytically active, and the most active one improved the activity of the template enzyme by 4 orders of magnitude. The computational details and parameter optimization of the sequence pairing step of the LoopGrafter workflow are revealed. The optimized and experimentally validated loop grafting workflow available as a fully automated Web server represents a powerful approach for engineering catalytically efficient enzymes by modification of protein dynamics.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

ACS Catalysis

ISSN

2155-5435

e-ISSN

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Volume of the periodical

15

Issue of the periodical within the volume

4

Country of publishing house

US - UNITED STATES

Number of pages

14

Pages from-to

3391-3404

UT code for WoS article

001418177600001

EID of the result in the Scopus database

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