Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP
I Protein

J 2024

Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein

SUBADINI, Suchismita; Krishnendu BERA; Devi Prasanna BEHERA; Jozef HRITZ; Harekrushna SAHOO et. al.

Základní údaje

Originální název

Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein

Autoři

SUBADINI, Suchismita; Krishnendu BERA; Devi Prasanna BEHERA; Jozef HRITZ a Harekrushna SAHOO

Vydání

Luminescence, HOBOKEN, Wiley, 2024, 1522-7235

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.000

Kód RIV

RIV/00216224:14310/24:00138215

Organizační jednotka

Přírodovědecká fakulta

DOI

https://doi.org/10.1002/bio.4924

UT WoS

001369889900001

EID Scopus

2-s2.0-85211138321

Klíčová slova anglicky

excluded volume effect; macromolecular crowder; PEG; protein refolding kinetics

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 3. 1. 2025 11:10, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Návaznosti

MUNI/G/1002/2021, interní kód MU

Název: Insight into CAIX structure and function and design of selective inhibitors as potential anti-cancer drugs (Akronym: CAIX-target)

Investor: Masarykova univerzita, Insight into CAIX structure and function and design of selective inhibitors as potential anti-cancer drugs, INTERDISCIPLINARY - Mezioborové výzkumné projekty

90254, velká výzkumná infrastruktura

Název: e-INFRA CZ II

Citovat

SUBADINI, Suchismita; Krishnendu BERA; Devi Prasanna BEHERA; Jozef HRITZ a Harekrushna SAHOO. Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein. Luminescence. HOBOKEN: Wiley, 2024, roč. 39, č. 12, s. 1-10. ISSN 1522-7235. Dostupné z: https://doi.org/10.1002/bio.4924.

@article{2464287,
   author = {Subadini, Suchismita and Bera, Krishnendu and Behera, Devi Prasanna and Hritz, Jozef and Sahoo, Harekrushna},
   article_location = {HOBOKEN},
   article_number = {12},
   doi = {https://doi.org/10.1002/bio.4924},
   keywords = {excluded volume effect; macromolecular crowder; PEG; protein refolding kinetics},
   language = {eng},
   issn = {1522-7235},
   journal = {Luminescence},
   title = {Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein},
   url = {https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4924},
   volume = {39},
   year = {2024}
}

TY  - JOUR
ID  - 2464287
AU  - Subadini, Suchismita - Bera, Krishnendu - Behera, Devi Prasanna - Hritz, Jozef - Sahoo, Harekrushna
PY  - 2024
TI  - Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein
JF  - Luminescence
VL  - 39
IS  - 12
SP  - 1-10
EP  - 1-10
PB  - Wiley
SN  - 15227235
KW  - excluded volume effect
KW  - macromolecular crowder
KW  - PEG
KW  - protein refolding kinetics
UR  - https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bio.4924
N2  - 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.
ER  -

SUBADINI, Suchismita; Krishnendu BERA; Devi Prasanna BEHERA; Jozef HRITZ a Harekrushna SAHOO. Polyethylene Glycol-Based Refolding Kinetic Modulation of CRABP I Protein. \textit{Luminescence}. HOBOKEN: Wiley, 2024, roč.~39, č.~12, s.~1-10. ISSN~1522-7235. Dostupné z: https://doi.org/10.1002/bio.4924.

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