VUV irradiation effects on proteins in high-flux synchrotron radiation circular dichroism spectroscopy

Synchrotron radiation circular dichroism (SRCD) spectroscopy is emerging as an important new tool in structural molecular biology. Previously we had shown that in lower‐flux SRCD instruments, such as UV1 at ISA and beamline 3.1 at the SRS, vacuum ultraviolet (VUV) radiation damage to proteins was no...

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Bibliographic Details
Published in:Journal of synchrotron radiation 2005-07, Vol.12 (4), p.517-523
Main Authors: Wien, F., Miles, A. J., Lees, J. G., Vrønning Hoffmann, S., Wallace, B. A.
Format: Article
Language:English
Subjects:
BASIC BIOLOGICAL SCIENCES
BIOCHEMISTRY
calibration
Circular Dichroism - methods
circular dichroism spectroscopy
Crystallography - methods
DICHROISM
Dose-Response Relationship, Radiation
IRRADIATION
membrane protein
national synchrotron light source
PARTICLE ACCELERATORS
Protein Conformation - radiation effects
protein denaturation
Protein Denaturation - radiation effects
protein structure
PROTEINS
Proteins - analysis
Proteins - chemistry
Proteins - radiation effects
Radiation Dosage
secondary structure
Spectrophotometry, Ultraviolet - methods
SPECTROSCOPY
structural genomics
SYNCHROTRON RADIATION
synchrotron radiation circular dichroism spectroscopy
Synchrotrons
Ultraviolet Rays
vacuum ultraviolet
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Summary:Synchrotron radiation circular dichroism (SRCD) spectroscopy is emerging as an important new tool in structural molecular biology. Previously we had shown that in lower‐flux SRCD instruments, such as UV1 at ISA and beamline 3.1 at the SRS, vacuum ultraviolet (VUV) radiation damage to proteins was not evident after exposure over a period of hours. No effects were detected in either the protein primary or the secondary structures. However, with the development of high‐flux beamlines, such as CD12 at the SRS, this issue has been revisited because of changes observed in the SRCD spectra of consecutive scans of protein samples obtained on this high‐flux beamline. Experiments have been designed to distinguish between two different possible mechanisms: (i) photoionization causing free radicals or secondary electrons producing degradation of the protein, and (ii) local heating of the sample resulting in protein denaturation. The latter appears to be the principal source of the signal deterioration.
ISSN:1600-5775
0909-0495
0021-9258
1600-5775
1083-351X
DOI:10.1107/S0909049505006953