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Free radicals and ROS induce protein denaturation by UV photostability assay

DOI: 10.3390/ijms22126512 DOI Help

Authors: Paolo Ruzza (CNR (ICB-CNR)) , Claudia Honisch (CNR (ICB-CNR); University of Padua) , Rohanah Hussain (Diamond Light Source) , Giuliano Siligardi (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: International Journal Of Molecular Sciences , VOL 22

State: Published (Approved)
Published: June 2021
Diamond Proposal Number(s): 13661 , 14857 , 15931 , 16643 , 17413

Open Access Open Access

Abstract: Oxidative stress, photo-oxidation, and photosensitizers are activated by UV irradiation and are affecting the photo-stability of proteins. Understanding the mechanisms that govern protein photo-stability is essential for its control enabling enhancement or reduction. Currently, two major mechanisms for protein denaturation induced by UV irradiation are available: one generated by the local heating of water molecules bound to the proteins and the other by the formation of reactive free radicals. To discriminate which is the likely or dominant mechanism we have studied the effects of thermal and UV denaturation of aqueous protein solutions with and without DHR-123 as fluorogenic probe using circular dichroism (CD), synchrotron radiation circular dichroism (SRCD), and fluorescence spectroscopies. The results indicated that the mechanism of protein denaturation induced by VUV and far-UV irradiation were mediated by the formation of reactive free radicals (FR) and reactive oxygen species (ROS). The development at Diamond B23 beamline for SRCD of a novel protein UV photo-stability assay based on consecutive repeated CD measurements in the far-UV (180–250 nm) region has been successfully used to assess and characterize the photo-stability of protein formulations and ligand binding interactions, in particular for ligand molecules devoid of significant UV absorption.

Journal Keywords: VUV and far-UV irradiation; protein photo-stability; synchrotron radiation circular dichroism; reactive oxygen species; UV protein denaturation assay

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: B23-Circular Dichroism

Added On: 21/06/2021 10:38

2021_Free Radicals and ROS Induce Protein Denaturation by UV Photostability Assay_Int J Mol Sci_Honisch et al_2021.pdf

Discipline Tags:

Life Sciences & Biotech Chemistry Biochemistry

Technical Tags:

Spectroscopy Circular Dichroism (CD)