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J Magn Reson. 2020 Feb;311:106680. doi: 10.1016/j.jmr.2019.106680. Epub 2019 Dec 23.

Sensitivity boosts by the CPMAS CryoProbe for challenging biological assemblies.

Author information

1
Bruker Biospin Corporation, Fällanden, Switzerland. Electronic address: Alia.Hassan@bruker.com.
2
Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, United States.
3
Bruker Biospin Corporation, 15 Fortune Drive, Billerica, MA, United States.
4
Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, United States; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States.
5
Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, United States.
6
CNRS, CBMN, UMR5248, University of Bordeaux, F-33600 Pessac, France.
7
Bruker Biospin Corporation, Fällanden, Switzerland.
8
Bruker Biospin Corporation, Fällanden, Switzerland. Electronic address: barbara.perrone@bruker.com.
9
Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States; Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Ave., Pittsburgh, PA, United States. Electronic address: amg100@pitt.edu.
10
Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, United States; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States. Electronic address: tpolenov@udel.edu.

Abstract

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™). Such intensity enhancements permit recording 2D and 3D experiments that are otherwise time-prohibitive, such as 2D 15N-15N proton-driven spin diffusion and 15N-13C 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.

KEYWORDS:

Biological assemblies; CryoProbe; MAS NMR; Magic angle spinning

PMID:
31951864
DOI:
10.1016/j.jmr.2019.106680

Conflict of interest statement

Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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