Publication details

Polyethylene glycol perturbs the unfolding of CRABP I: A correlation between experimental and theoretical approach

Investor logo
Investor logo
Authors

SUBADINI Suchismita BERA Krishnendu HRITZ Jozef SAHOO Harekrushna

Year of publication 2021
Type Article in Periodical
Magazine / Source Colloids and Surfaces B: Biointerfaces
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://doi.org/10.1016/j.colsurfb.2021.111696
Doi http://dx.doi.org/10.1016/j.colsurfb.2021.111696
Keywords PEG; protein stabilizer; Crowding agent; excluded volume; protein unfolding
Description The importance of macromolecules paves the way towards a detailed molecular level investigation as all most all cellular processes occurring at the interior of cells in the form of proteins, enzymes, and other biological molecules are significantly affected because of their crowding. Thus, exploring the role of crowding environment on the denaturation and renaturation kinetics of protein molecules is of great importance. Here, we have employed CRABP I (cellular retinoic acid binding protein I), as a model protein along with different molecular weights of Polyethylene glycol (PEG) as molecular crowders. The experimental evaluations were done by accessing the protein secondary structure analysis using circular dichroism (CD) spectroscopy and unfolding kinetics using intrinsic fluorescence of CRABP I at 37?°C to mimic the in vivo crowding environment. The unfolding kinetics results indicated that both PEG 2000 and PEG 4000 act as stabilizers by retarding the unfolding kinetic rates. Both kinetic and stability outcomes presented the importance of crowding environment on stability and kinetics of CRABP I. The molecular dynamics (MD) studies revealed that thirteen PEG 2000 molecules assembled during the 500?ns simulation, which increases the stability and percentage of ß-sheet. The experimental findings were well supported by the molecular dynamics simulation results.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.

More info