Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F24%3A00138215" target="_blank" >RIV/00216224:14310/24:00138215 - isvavai.cz</a>

Result on the web

<a href="https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4924" target="_blank" >https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4924</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/bio.4924" target="_blank" >10.1002/bio.4924</a>

Result language

angličtina

Original language name

Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein

Original language description

Crowding environment has a significant impact on the folding and stability of protein in biological systems. In this work, we have used four different sizes of a molecular crowder, polyethylene glycol (PEG), to analyze the unfolding and refolding kinetics of an iLBP protein, CRABP I, using urea as chemical denaturant. In general, the stability of the native state of the protein is boosted by the presence of crowding agents in the solution. However, our findings show that not only the type of crowder but also the crowder size played a key role in the effects of excluded volume. In case of lower molecular weight of PEG (M.W. 400), even at 200 g/L concentration, only the viscosity effect is observed, whereas for higher molecular weight of PEG (M.W. 1000), both the viscosity effect and excluded volume effect are noticed, and even at a higher concentration (200 g/L) of PEG 1000, the excluded volume predominates over the viscosity effect. Using the transition state theory, we were also able to determine the free energies of activation for the unfolding and refolding studies from their respective rate constants. Additionally, MD simulation studies provide strong support for our experimental observation. Analysis of secondary structure propensity (SSP) reveals a marked decline in the presence of structural elements (beta-sheet, beta-bridge, turn, and alpha-helix) from 81% to 43% over the 1 mu s time scale unfolding MD simulation under 8 M urea conditions. Conversely, in a 200 ns refolding simulation, the rate of refolding notably increases at a concentration of 200 g/L PEG 1000.

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

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2024

Confidentiality

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

Name of the periodical

Luminescence

ISSN

1522-7235

e-ISSN

—

Volume of the periodical

39

Issue of the periodical within the volume

12

Country of publishing house

US - UNITED STATES

Number of pages

10

Pages from-to

1-10

UT code for WoS article

001369889900001

EID of the result in the Scopus database

2-s2.0-85211138321