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淀粉样蛋白与膜相互作用的固态核磁共振研究

Solid-state NMR of amyloid membrane interactions.

作者信息

Gehman John D, Separovic Frances

机构信息

School of Chemistry, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, VIC, Australia.

出版信息

Methods Mol Biol. 2011;752:165-77. doi: 10.1007/978-1-60327-223-0_11.

Abstract

Solid-state NMR pulse sequences often feature fewer pulses and delays than the more common solution NMR experiments. This ostensible simplicity, however, belies the care with which experimental parameters must be determined, as solid-state NMR can be much less forgiving of improper experimental set-up. This is especially true of "semi-solid" samples, such as the phospholipid vesicles used to study membrane-associated peptides and proteins, which feature prominently in misfolding diseases. Protocols for the preparation of multilamellar vesicles for solid-state NMR studies of Aβ peptides are described, together with procedures for optimization of critical experimental parameters, such as spectral widths, delay times, and field strengths for (31)P, (2)H, and (13)C NMR spectroscopy.

摘要

与更常见的溶液核磁共振实验相比,固态核磁共振脉冲序列通常具有更少的脉冲和延迟。然而,这种表面上的简单掩盖了确定实验参数时所需的谨慎,因为固态核磁共振对不当的实验设置宽容度要低得多。对于“半固态”样品尤其如此,例如用于研究与膜相关的肽和蛋白质的磷脂囊泡,它们在错误折叠疾病中显著存在。本文描述了用于Aβ肽固态核磁共振研究的多层囊泡制备方案,以及用于优化关键实验参数的程序,如用于(31)P、(2)H和(13)C核磁共振光谱的光谱宽度、延迟时间和场强。

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