Protein fiber linear dichroism for structure determination and kinetics in a low-volume, low-wavelength couette flow cell

被引:62
作者
Dafforn, TR
Rajendra, J
Halsall, DJ
Serpell, LC
Rodger, A
机构
[1] Univ Manchester, Dept Biol Sci, Manchester M13 9PT, Lancs, England
[2] Univ Warwick, Dept Chem, Coventry CV4 7AL, W Midlands, England
[3] Univ Cambridge, Cambridge Inst Med Res, Dept Haematol, Cambridge CB2 2XY, England
[4] Univ Cambridge, Cambridge Inst Med Res, Dept Clin Biochem, Cambridge CB2 2XY, England
[5] MRC, Mol Biol Lab, Div Neurobiol, Cambridge CB2 2QH, England
基金
英国工程与自然科学研究理事会;
关键词
D O I
10.1016/S0006-3495(04)74116-8
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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 muL), low-wavelength (down to 180 nm), low-pathlength (100 mum), 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-beta-sheet amyloid fibers of the Alzheimer's derived protein Abeta and the long-chain assemblies of alpha(1)-antitrypsin polymers.
引用
收藏
页码:404 / 410
页数:7
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