Decoding the intricate network of molecular interactions of a hyperstable engineered biocatalyst

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F20%3A00073505" target="_blank" >RIV/00159816:_____/20:00073505 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/20:00117258

Result on the web

<a href="https://pubs.rsc.org/en/content/articlelanding/2020/SC/D0SC03367G#!divAbstract" target="_blank" >https://pubs.rsc.org/en/content/articlelanding/2020/SC/D0SC03367G#!divAbstract</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1039/d0sc03367g" target="_blank" >10.1039/d0sc03367g</a>

Result language

angličtina

Original language name

Decoding the intricate network of molecular interactions of a hyperstable engineered biocatalyst

Original language description

Computational design of protein catalysts with enhanced stabilities for use in research and enzyme technologies is a challenging task. Using force-field calculations and phylogenetic analysis, we previously designed the haloalkane dehalogenase DhaA115 which contains 11 mutations that confer upon it outstanding thermostability (T-m = 73.5 degrees C; Delta T-m &gt; 23 degrees C). An understanding of the structural basis of this hyperstabilization is required in order to develop computer algorithms and predictive tools. Here, we report X-ray structures of DhaA115 at 1.55 angstrom and 1.6 angstrom resolutions and their molecular dynamics trajectories, which unravel the intricate network of interactions that reinforce the aba-sandwich architecture. Unexpectedly, mutations toward bulky aromatic amino acids at the protein surface triggered long-distance (similar to 27 angstrom) backbone changes due to cooperative effects. These cooperative interactions produced an unprecedented double-lock system that: (i) induced backbone changes, (ii) closed the molecular gates to the active site, (iii) reduced the volumes of the main and slot access tunnels, and (iv) occluded the active site. Despite these spatial restrictions, experimental tracing of the access tunnels using krypton derivative crystals demonstrates that transport of ligands is still effective. Our findings highlight key thermostabilization effects and provide a structural basis for designing new thermostable protein catalysts.

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

10406 - Analytical 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

2020

Confidentiality

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

Name of the periodical

Chemical Science

ISSN

2041-6520

e-ISSN

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Volume of the periodical

11

Issue of the periodical within the volume

41

Country of publishing house

GB - UNITED KINGDOM

Number of pages

17

Pages from-to

11162-11178

UT code for WoS article

000582936200030

EID of the result in the Scopus database

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