ArticleInternational journal of molecular sciences2021
Free Radical Generation in Far-UV Synchrotron Radiation Circular Dichroism Assays-Protein and Buffer Composition Contribution.
Article in International journal of molecular sciences, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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5 citing papers in PubMed, 7 citations in OpenAlex.
- Enzyme-Ligand Interaction Monitored by Synchrotron Radiation Circular Dichroism.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Enhanced Protein Photo-Stability Analysis Using SRCD in the Presence of Phospholipid SUVs.Chemistry (Weinheim an der Bergstrasse, Germany) · 2025Article
- New Perspective on the Interaction Behavior Between Riboflavin and β Lactoglobulin-β Casein Complex by Biophysical Techniques.Cell biochemistry and biophysics · 2024Article
- Circular Dichroism Study of Orexin B under Oxidative Stress Conditions.Molecules (Basel, Switzerland) · 2023Article
- Analysis of aptamer-target binding and molecular mechanisms by thermofluorimetric analysis and molecular dynamics simulation.Frontiers in chemistry · 2023Article
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Authors and funding
4 authors at 3 institutions in 2 countries.
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Abstract
A useful tool to analyze the ligands and/or environmental contribution to protein stability is represented by the Synchrotron Radiation Circular Dichroism UV-denaturation assay that consists in the acquisition of several consecutive repeated far-UV SRCD spectra. Recently we demonstrated that the prevailing mechanism of this denaturation involves the generation of free radicals and reactive oxygen species (ROS). In this work, we analyzed the effect of buffering agents commonly used in spectroscopic measurements, including MOPS (3-(N-morpholino) propanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TRIS-HCl (tris-hydroxymethil aminomethane hydrochloride), and phosphate, on the efficiency of protein denaturation caused by exposure to UV radiation. Fluorescence experiments confirmed the presence of ROS and were used to determine the rate of ROS generation. Our results indicate that the efficiency of the denaturation process is strongly influenced by the buffer composition with MOPS and HEPES acting also as scavengers and that the presence of proteins itself influenced the ROS formation rate.
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