Elongation rate and average length of amyloid fibrils in solution using isotope-labelled small-angle neutron scattering.
Elongation rate and average length of amyloid fibrils in solution using isotope-labelled small-angle neutron scattering.
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DOI：
10.1039/d1cb00001b
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发表时间：
2021-08-05
影响因子：
4.1
通讯作者：
Squires AM
中科院分区：
文献类型：
--
作者：
Eves BJ;Doutch JJ;Terry AE;Yin H;Moulin M;Haertlein M;Forsyth VT;Flagmeier P;Knowles TPJ;Dias DM;Lotze G;Seddon AM;Squires AM
我们展示了一种解决方案的方法，允许伸长率和平均纤维长度的组装淀粉样蛋白原纤维进行估计。该方法包括在种子生长的初始阶段采集实时中子散射数据，使用对比度匹配缓冲区使种子有效地对中子不可见。作为氘代单体添加到种子，标记的增长结束引起的散射模式，我们的模型作为圆柱体的长度随时间的增加给出了伸长率。此外，信号的绝对强度可用于确定每单位体积的生长端的数量，这进而提供种子长度的估计。在伸长过程中，末端的数量没有显著变化，这表明与预先存在的原纤维末端的生长相比，任何自发或二次成核都不显著，此外还提供了该技术的内部验证方法。我们在室温下使用2.5 mg ml−1氘代单体中的1.2 mg ml−1种子进行α突触核蛋白原纤维的初始生长实验，得到的延伸率为6.3 ± 0.5 μ min−1，平均种子长度估计为4.2 ± 1.3 μm。我们证明了一种解决方案的方法，允许伸长率和淀粉样纤维的平均长度被独立地确定。
We demonstrate a solution method that allows both elongation rate and average fibril length of assembling amyloid fibrils to be estimated. The approach involves acquisition of real-time neutron scattering data during the initial stages of seeded growth, using contrast matched buffer to make the seeds effectively invisible to neutrons. As deuterated monomers add on to the seeds, the labelled growing ends give rise to scattering patterns that we model as cylinders whose increase in length with time gives an elongation rate. In addition, the absolute intensity of the signal can be used to determine the number of growing ends per unit volume, which in turn provides an estimate of seed length. The number of ends did not change significantly during elongation, demonstrating that any spontaneous or secondary nucleation was not significant compared with growth on the ends of pre-existing fibrils, and in addition providing a method of internal validation for the technique. Our experiments on initial growth of alpha synuclein fibrils using 1.2 mg ml−1 seeds in 2.5 mg ml−1 deuterated monomer at room temperature gave an elongation rate of 6.3 ± 0.5 Å min−1, and an average seed length estimate of 4.2 ± 1.3 μm. We demonstrate a solution method that allows both elongation rate and average length of amyloid fibrils to be independently determined.
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影响因子：
4.8
作者：
Green, JD;Goldsbury, C;Aebi, U
通讯作者：
Aebi, U
影响因子：
5
作者：
Hellstrand, Erik;Grey, Marie;Sparr, Emma
通讯作者：
Sparr, Emma
DOI：
10.1016/j.nima.2014.07.029
发表时间：
2014-11-11
影响因子：
1.4
作者：
Arnold, O.;Bilheux, J. C.;Zikoysky, J.
通讯作者：
Zikoysky, J.
DOI：
10.1073/pnas.96.7.3688
发表时间：
1999-03-30
影响因子：
11.1
作者：
Kowalewski, T;Holtzman, DM
通讯作者：
Holtzman, DM
DOI：
10.1039/c4cp05563b
发表时间：
2015-03-28
期刊：
Physical chemistry chemical physics : PCCP
影响因子：
--
作者：
Arosio P;Knowles TP;Linse S
通讯作者：
Linse S