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Protein fiber linear dichroism for structure determination and kinetics in a low-volume, low-wavelength couette flow cell
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Dafforn, Tim, Rajendra, Jacindra, Halsall, David J., Serpell, Louise C. and Rodger, Alison (2004) Protein fiber linear dichroism for structure determination and kinetics in a low-volume, low-wavelength couette flow cell. Biophysical Journal, Vol.86 (No.1). pp. 404-410. doi:10.1016/S0006-3495(04)74116-8 ISSN 0006-3495.
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Official URL: http://dx.doi.org/10.1016/S0006-3495(04)74116-8
Abstract
High-resolution structure determination of soluble globular proteins relies heavily on x-ray crystallography techniques. Such an approach is often ineffective for investigations into the structure of fibrous proteins as these proteins generally do not crystallize. Thus investigations into fibrous protein structure have relied on less direct methods such as x-ray fiber diffraction and circular dichroism. Ultraviolet linear dichroism has the potential to provide additional information on the structure of such biomolecular systems. However, existing systems are not optimized for the requirements of fibrous proteins. We have designed and built a low-volume (200 μL), low-wavelength (down to 180 nm), low-pathlength (100 μm), high-alignment flow-alignment system (couette) to perform ultraviolet linear dichroism studies on the fibers formed by a range of biomolecules. The apparatus has been tested using a number of proteins for which longer wavelength linear dichroism spectra had already been measured. The new couette cell has also been used to obtain data on two medically important protein fibers, the all-β-sheet amyloid fibers of the Alzheimer's derived protein Aβ and the long-chain assemblies of α1-antitrypsin polymers.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||
Library of Congress Subject Headings (LCSH): | X-ray crystallography -- Technique, Globular proteins, Linear dichroism, Laminar flow, Protein binding | ||||
Journal or Publication Title: | Biophysical Journal | ||||
Publisher: | Biophysical Society | ||||
ISSN: | 0006-3495 | ||||
Official Date: | January 2004 | ||||
Dates: |
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Volume: | Vol.86 | ||||
Number: | No.1 | ||||
Page Range: | pp. 404-410 | ||||
DOI: | 10.1016/S0006-3495(04)74116-8 | ||||
Status: | Peer Reviewed | ||||
Access rights to Published version: | Open Access (Creative Commons) | ||||
Funder: | Engineering and Physical Sciences Research Council (EPSRC), Wellcome Trust (London, England), Medical Research Council (Great Britain) (MRC) | ||||
Grant number: | GR/M91105 (EPSRC), GR/R40869/01 (EPSRC) |
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