Substrate inhibition by the blockage of product release and its control by tunnel engineering

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F21%3A00075125" target="_blank" >RIV/00159816:_____/21:00075125 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/21:00122273

Result on the web

<a href="https://pubs.rsc.org/en/content/articlelanding/2021/CB/D0CB00171F" target="_blank" >https://pubs.rsc.org/en/content/articlelanding/2021/CB/D0CB00171F</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1039/d0cb00171f" target="_blank" >10.1039/d0cb00171f</a>

Result language

angličtina

Original language name

Substrate inhibition by the blockage of product release and its control by tunnel engineering

Original language description

Substrate inhibition is the most common deviation from Michaelis-Menten kinetics, occurring in approximately 25% of known enzymes. It is generally attributed to the formation of an unproductive enzyme-substrate complex after the simultaneous binding of two or more substrate molecules to the active site. Here, we show that a single point mutation (L177W) in the haloalkane dehalogenase LinB causes strong substrate inhibition. Surprisingly, a global kinetic analysis suggested that this inhibition is caused by binding of the substrate to the enzyme-product complex. Molecular dynamics simulations clarified the details of this unusual mechanism of substrate inhibition: Markov state models indicated that the substrate prevents the exit of the halide product by direct blockage and/or restricting conformational flexibility. The contributions of three residues forming the possible substrate inhibition site (W140A, F143L and I211L) to the observed inhibition were studied by mutagenesis. An unusual synergy giving rise to high catalytic efficiency and reduced substrate inhibition was observed between residues L177W and I211L, which are located in different access tunnels of the protein. These results show that substrate inhibition can be caused by substrate binding to the enzyme-product complex and can be controlled rationally by targeted amino acid substitutions in enzyme access tunnels.

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

R - Projekt Ramcoveho programu EK

Publication year

2021

Confidentiality

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

Name of the periodical

RSC CHEMICAL BIOLOGY

ISSN

2633-0679

e-ISSN

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Volume of the periodical

2

Issue of the periodical within the volume

2

Country of publishing house

GB - UNITED KINGDOM

Number of pages

11

Pages from-to

645-655

UT code for WoS article

000641784900024

EID of the result in the Scopus database

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