Fraga, Hugo and Arnaud, Charles-Adrien and Gauto, Diego F. and Audin, Maxime and Kurauskas, Vilius and Macek, Pavel and Krichel, Carsten and Guan, Jia-Ying and Boisbouvier, Jerome and Sprangers, Remco and Breyton, Cecile and Schanda, Paul (2017) Solid-State NMR H-N-(C)-H and H-N-C-C 3D/4D Correlation Experiments for Resonance Assignment of Large Proteins. CHEMPHYSCHEM, 18 (19). pp. 2697-2703. ISSN 1439-4235, 1439-7641
Full text not available from this repository. (Request a copy)Abstract
Solid-state NMR spectroscopy can provide insight into protein structure and dynamics at the atomic level without inherent protein size limitations. However, a major hurdle to studying large proteins by solid-state NMR spectroscopy is related to spectral complexity and resonance overlap, which increase with molecular weight and severely hamper the assignment process. Here the use of two sets of experiments is shown to expand the tool kit of H-1-detected assignment approaches, which correlate a given amide pair either to the two adjacent CO-CA pairs (4D hCOCANH/hCOCAcoNH), or to the amide H-1 of the neighboring residue (3D HcocaNH/HcacoNH, which can be extended to 5D). The experiments are based on efficient coherence transfers between backbone atoms using INEPT transfers between carbons and cross-polarization for heteronuclear transfers. The utility of these experiments is exemplified with application to assemblies of deuterated, fully amide-protonated proteins from approximately 20 to 60kDa monomer, at magic-angle spinning (MAS) frequencies from approximately 40 to 55kHz. These experiments will also be applicable to protonated proteins at higher MAS frequencies. The resonance assignment of a domain within the 50.4kDa bacteriophageT5 tube protein pb6 is reported, and this is compared to NMR assignments of the isolated domain in solution. This comparison reveals contacts of this domain to the core of the polymeric tail tube assembly.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROTON-DETECTED NMR; ANGLE-SPINNING NMR; BACKBONE ASSIGNMENT; PERDEUTERATED PROTEINS; HIGH-RESOLUTION; ATOMIC MODEL; FAST MAS; 100 KHZ; SPECTROSCOPY; LINKING; NMR methods; NMR pulse sequences; phage proteins; protein assemblies; protein quality control machinery |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:19 |
| Last Modified: | 11 Feb 2019 15:26 |
| URI: | https://pred.uni-regensburg.de/id/eprint/2022 |
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