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

Kód výsledku v 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>

Nalezeny alternativní kódy

RIV/00216224:14310/20:00117258

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2020

Kód důvěrnosti údajů

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

Název periodika

Chemical Science

ISSN

2041-6520

e-ISSN

—

Svazek periodika

11

Číslo periodika v rámci svazku

41

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

17

Strana od-do

11162-11178

Kód UT WoS článku

000582936200030

EID výsledku v databázi Scopus

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