UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD
Light irradiation with high photon flux in the vacuum and far-UV region is known to denature the conformation of biopolymers. Measures are in place at Diamond Light Source B23 beamline for Synchrotron Radiation Circular Dichroism (SRCD) to control and make this effect negligible. However, UV denatur...
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MDPI AG
2018-07-01
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Series: | Molecules |
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Online Access: | http://www.mdpi.com/1420-3049/23/8/1906 |
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author | Rohanah Hussain Edoardo Longo Giuliano Siligardi |
author_facet | Rohanah Hussain Edoardo Longo Giuliano Siligardi |
author_sort | Rohanah Hussain |
collection | DOAJ |
description | Light irradiation with high photon flux in the vacuum and far-UV region is known to denature the conformation of biopolymers. Measures are in place at Diamond Light Source B23 beamline for Synchrotron Radiation Circular Dichroism (SRCD) to control and make this effect negligible. However, UV denaturation of proteins can also be exploited as a novel method for assessing biopolymer photostability as well as ligand-binding interactions. Usually, host–ligand binding interactions can be assessed monitoring CD changes of the host biopolymer upon ligand addition. The novel method of identifying ligand binding monitoring the change of relative rate of UV denaturation using SRCD is especially important when there are very little or insignificant secondary structure changes of the host protein upon ligand binding. The temperature study, another method used to determine molecular interactions, can often be inconclusive when the thermal effect associated with the displacement of the bound solvent molecules by the ligand is also small, making the determination of the binding interaction inconclusive. Herein we present a review on the UV-denaturation assay as a novel method to determine the relative photostability of protein formulations as well as the screening of ligand-binding interactions using the high photon flux Diamond B23 beamline for SRCD. |
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id | doaj.art-5382a849634b4086ae71d8795d468a11 |
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issn | 1420-3049 |
language | English |
last_indexed | 2024-12-21T22:21:32Z |
publishDate | 2018-07-01 |
publisher | MDPI AG |
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series | Molecules |
spelling | doaj.art-5382a849634b4086ae71d8795d468a112022-12-21T18:48:19ZengMDPI AGMolecules1420-30492018-07-01238190610.3390/molecules23081906molecules23081906UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCDRohanah Hussain0Edoardo Longo1Giuliano Siligardi2B23 Beamline, Diamond Light Source, Harwell Science Innovation Campus, Chilton, Didcot OX11 0DE, UKB23 Beamline, Diamond Light Source, Harwell Science Innovation Campus, Chilton, Didcot OX11 0DE, UKB23 Beamline, Diamond Light Source, Harwell Science Innovation Campus, Chilton, Didcot OX11 0DE, UKLight irradiation with high photon flux in the vacuum and far-UV region is known to denature the conformation of biopolymers. Measures are in place at Diamond Light Source B23 beamline for Synchrotron Radiation Circular Dichroism (SRCD) to control and make this effect negligible. However, UV denaturation of proteins can also be exploited as a novel method for assessing biopolymer photostability as well as ligand-binding interactions. Usually, host–ligand binding interactions can be assessed monitoring CD changes of the host biopolymer upon ligand addition. The novel method of identifying ligand binding monitoring the change of relative rate of UV denaturation using SRCD is especially important when there are very little or insignificant secondary structure changes of the host protein upon ligand binding. The temperature study, another method used to determine molecular interactions, can often be inconclusive when the thermal effect associated with the displacement of the bound solvent molecules by the ligand is also small, making the determination of the binding interaction inconclusive. Herein we present a review on the UV-denaturation assay as a novel method to determine the relative photostability of protein formulations as well as the screening of ligand-binding interactions using the high photon flux Diamond B23 beamline for SRCD.http://www.mdpi.com/1420-3049/23/8/1906circular dichroismligand bindinghigh photon fluxprotein stabilitysynchrotron radiationSRCDvacuum UV |
spellingShingle | Rohanah Hussain Edoardo Longo Giuliano Siligardi UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD Molecules circular dichroism ligand binding high photon flux protein stability synchrotron radiation SRCD vacuum UV |
title | UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD |
title_full | UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD |
title_fullStr | UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD |
title_full_unstemmed | UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD |
title_short | UV-Denaturation Assay to Assess Protein Photostability and Ligand-Binding Interactions Using the High Photon Flux of Diamond B23 Beamline for SRCD |
title_sort | uv denaturation assay to assess protein photostability and ligand binding interactions using the high photon flux of diamond b23 beamline for srcd |
topic | circular dichroism ligand binding high photon flux protein stability synchrotron radiation SRCD vacuum UV |
url | http://www.mdpi.com/1420-3049/23/8/1906 |
work_keys_str_mv | AT rohanahhussain uvdenaturationassaytoassessproteinphotostabilityandligandbindinginteractionsusingthehighphotonfluxofdiamondb23beamlineforsrcd AT edoardolongo uvdenaturationassaytoassessproteinphotostabilityandligandbindinginteractionsusingthehighphotonfluxofdiamondb23beamlineforsrcd AT giulianosiligardi uvdenaturationassaytoassessproteinphotostabilityandligandbindinginteractionsusingthehighphotonfluxofdiamondb23beamlineforsrcd |