Decoding Protein Stabilization: Impact on Aggregation, Solubility, and Unfolding Mechanisms

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F25%3A00082420" target="_blank" >RIV/00159816:_____/25:00082420 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14310/25:00142062

Výsledek na webu

<a href="https://pubs.acs.org/doi/10.1021/acs.jcim.5c00611" target="_blank" >https://pubs.acs.org/doi/10.1021/acs.jcim.5c00611</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acs.jcim.5c00611" target="_blank" >10.1021/acs.jcim.5c00611</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Decoding Protein Stabilization: Impact on Aggregation, Solubility, and Unfolding Mechanisms

Popis výsledku v původním jazyce

Modern computational tools can predict the mutational effects on protein stability, sometimes at the expense of activity or solubility. Here, we investigate two homologous computationally stabilized haloalkane dehalogenases: (i) the soluble thermostable DhaA115 (T m app = 74 degrees C) and (ii) the poorly soluble and aggregating thermostable LinB116 (T m app = 65 degrees C), together with their respective wild-type variants. The intriguing difference in the solubility of these highly homologous proteins has remained unexplained for three decades. We combined experimental and in-silico techniques and examined the effects of stabilization on solubility and aggregation propensity. A detailed analysis of the unfolding mechanisms in the context of aggregation explained the negative consequences of stabilization observed in LinB116. With the aid of molecular dynamics simulations, we identified regions exposed during the unfolding of LinB116 that were later found to exhibit aggregation propensity. Our analysis identified cryptic aggregation-prone regions and increased surface hydrophobicity as key factors contributing to the reduced solubility of LinB116. This study reveals novel molecular mechanisms of unfolding for hyperstabilized dehalogenases and highlights the importance of contextual information in protein engineering to avoid the negative effects of stabilizing mutations on protein solubility.

Název v anglickém jazyce

Decoding Protein Stabilization: Impact on Aggregation, Solubility, and Unfolding Mechanisms

Popis výsledku anglicky

Modern computational tools can predict the mutational effects on protein stability, sometimes at the expense of activity or solubility. Here, we investigate two homologous computationally stabilized haloalkane dehalogenases: (i) the soluble thermostable DhaA115 (T m app = 74 degrees C) and (ii) the poorly soluble and aggregating thermostable LinB116 (T m app = 65 degrees C), together with their respective wild-type variants. The intriguing difference in the solubility of these highly homologous proteins has remained unexplained for three decades. We combined experimental and in-silico techniques and examined the effects of stabilization on solubility and aggregation propensity. A detailed analysis of the unfolding mechanisms in the context of aggregation explained the negative consequences of stabilization observed in LinB116. With the aid of molecular dynamics simulations, we identified regions exposed during the unfolding of LinB116 that were later found to exhibit aggregation propensity. Our analysis identified cryptic aggregation-prone regions and increased surface hydrophobicity as key factors contributing to the reduced solubility of LinB116. This study reveals novel molecular mechanisms of unfolding for hyperstabilized dehalogenases and highlights the importance of contextual information in protein engineering to avoid the negative effects of stabilizing mutations on protein solubility.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

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

2025

Kód důvěrnosti údajů

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

Název periodika

Journal of Chemical Information and Modeling

ISSN

1549-9596

e-ISSN

1549-960X

Svazek periodika

65

Číslo periodika v rámci svazku

16

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

14

Strana od-do

8688-8701

Kód UT WoS článku

001545173800001

EID výsledku v databázi Scopus

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