Sensitivity boosts by the CPMAS CryoProbe for challenging biological assemblies
Sensitivity boosts by the CPMAS CryoProbe for challenging biological assemblies
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DOI：
10.1016/j.jmr.2019.106680
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发表时间：
2020-02-01
影响因子：
2.2
通讯作者：
Polenova, Tatyana
中科院分区：
文献类型：
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作者：
Hassan, Alia;Quinn, Caitlin M.;Polenova, Tatyana
尽管MAS NMR硬件和实验方法取得了突破，但灵敏度仍然是大型复杂生物系统的主要挑战。在这里，我们报告说，3-4倍的灵敏度可以在异核检测实验中获得，使用一种新的HCN CPMAS探针，其中样品线圈和电子设备在低温下工作，而样品保持在环境温度（BioSolids CryoProbe（TM））。这种强度增强允许记录否则时间限制的2D和3D实验，例如2D N-15-N-15质子驱动的自旋扩散和N-15-C-13双交叉极化到天然丰度碳实验。CPMAS CryoProbe为基础的实验的好处是说明驱动蛋白Kif 5 b与微管，HIV-1衣壳蛋白组件，和人类Y145 Stop和真菌HET-s朊病毒蛋白质的原纤维的组件要求系统的常规MAS固态NMR和优良的参考系统的光谱质量。我们设想，这种探针技术将有利于广泛的应用，特别是对生物系统的固有灵敏度低，在生理温度下的痛苦。(C)2019爱思唯尔公司All rights reserved.
Despite breakthroughs in MAS NMR hardware and experimental methodologies, sensitivity remains a major challenge for large and complex biological systems. Here, we report that 3-4 fold higher sensitivities can be obtained in heteronuclear-detected experiments, using a novel HCN CPMAS probe, where the sample coil and the electronics operate at cryogenic temperatures, while the sample is maintained at ambient temperatures (BioSolids CryoProbe (TM)). Such intensity enhancements permit recording 2D and 3D experiments that are otherwise time-prohibitive, such as 2D N-15-N-15 proton-driven spin diffusion and N-15-C-13 double cross polarization to natural abundance carbon experiments. The benefits of CPMAS CryoProbe-based experiments are illustrated for assemblies of kinesin Kif5b with microtubules, HIV-1 capsid protein assemblies, and fibrils of human Y145Stop and fungal HET-s prion proteins demanding systems for conventional MAS solid-state NMR and excellent reference systems in terms of spectral quality. We envision that this probe technology will be beneficial for a wide range of applications, especially for biological systems suffering from low intrinsic sensitivity and at physiological temperatures. (C) 2019 Elsevier Inc. All rights reserved.