Decoding Protein Stabilization: Impact on Aggregation, Solubility, and Unfolding Mechanisms
The result's identifiers
Result code in 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>
Alternative codes found
RIV/00216224:14310/25:00142062
Result on the web
<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>
Alternative languages
Result language
angličtina
Original language name
Decoding Protein Stabilization: Impact on Aggregation, Solubility, and Unfolding Mechanisms
Original language description
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.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - 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
Result continuities
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)
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Chemical Information and Modeling
ISSN
1549-9596
e-ISSN
1549-960X
Volume of the periodical
65
Issue of the periodical within the volume
16
Country of publishing house
US - UNITED STATES
Number of pages
14
Pages from-to
8688-8701
UT code for WoS article
001545173800001
EID of the result in the Scopus database
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