Impact of the access tunnel engineering on catalysis is strictly ligand-specific

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F18%3A00100915" target="_blank" >RIV/00216224:14310/18:00100915 - isvavai.cz</a>

Alternative codes found

RIV/00159816:_____/18:00068672

Result on the web

<a href="https://doi.org/10.1111/febs.14418" target="_blank" >https://doi.org/10.1111/febs.14418</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1111/febs.14418" target="_blank" >10.1111/febs.14418</a>

Result language

angličtina

Original language name

Impact of the access tunnel engineering on catalysis is strictly ligand-specific

Original language description

The traditional way of rationally engineering enzymes to change their biocatalytic properties utilizes the modifications of their active sites. Another emerging approach is the engineering of structural features involved in the exchange of ligands between buried active sites and the surrounding solvent. However, surprisingly little is known about the effects of mutations that alter the access tunnels on the enzymes’ catalytic properties, and how these tunnels should be redesigned to allow fast passage of cognate substrates and products. Thus, we have systematically studied the effects of single-point mutations in a tunnel-lining residue of a haloalkane dehalogenase on the binding kinetics and catalytic conversion of both linear and branched haloalkanes. The hotspot residue Y176 was identified using computer simulations and randomized through saturation mutagenesis, and the resulting variants were screened for shifts in binding rates. Strikingly, opposite effects of the substituted residues on the catalytic efficiency toward linear and branched substrates were observed, which was found to be due to substrate-specific requirements in the critical steps of the respective catalytic cycles. We conclude that not only the catalytic sites, but also the access pathways must be tailored specifically for each individual ligand, which is a new paradigm in protein engineering and de novo protein design. A rational approach is proposed here to address more effectively the task of designing ligand-specific tunnels using computational tools.

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

10401 - Organic chemistry

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

2018

Confidentiality

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

Name of the periodical

the FEBS Journal

ISSN

1742-464X

e-ISSN

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Volume of the periodical

285/2018

Issue of the periodical within the volume

8

Country of publishing house

GB - UNITED KINGDOM

Number of pages

21

Pages from-to

1456-1476

UT code for WoS article

000430691000007

EID of the result in the Scopus database

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