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Protein Fiber Linear Dichroism for Structure Determination and Kinetics in a Low-Volume, Low-Wavelength Couette Flow Cell

journal contribution
posted on 2023-06-07, 19:53 authored by Timothy R Dafforn, Jacindra Rajendra, David J Halsall, Louise SerpellLouise Serpell, Alison Rodger
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-ß-sheet amyloid fibers of the Alzheimer's derived protein Aß and the long-chain assemblies of a1-antitrypsin polymers.

History

Publication status

  • Published

Journal

Biophysical Journal

ISSN

0006-3495

Issue

1

Volume

86

Page range

404-410

Pages

7.0

Department affiliated with

  • Biochemistry Publications

Full text available

  • No

Peer reviewed?

  • Yes

Legacy Posted Date

2012-02-06

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