期刊
SOLID STATE NUCLEAR MAGNETIC RESONANCE
卷 87, 期 -, 页码 117-125出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ssnmr.2017.07.001
关键词
MAS NMR; Protein assemblies; Proton-detected MAS NMR correlations; HIV-1 capsid protein
资金
- National Institutes of Health (NIH) [P50 GM082251, P30GM110758]
- National Science Foundation (NSF) [CHE0959496]
The recent breakthroughs in NMR probe technologies resulted in the development of MAS NMR probes with rotation frequencies exceeding 100 kHz. Herein, we explore dramatic increases in sensitivity and resolution observed at MAS frequencies of 110-111 kHz in a novel 0.7 mm HCND probe that enable structural analysis of fully protonated biological systems. Proton-detected 2D and 3D correlation spectroscopy under such conditions requires only 0.1-0.5 mg of sample and a fraction of time compared to conventional C-13-detected experiments. We discuss the performance of several proton-and heteronuclear- (C-13-, N-15-) based correlation experiments in terms of sensitivity and resolution, using a model microcrystalline fMLF tripeptide. We demonstrate the applications of ultrafast MAS to a large, fully protonated protein assembly of the 231-residue HIV-1 CA capsid protein. Resonance assignments of protons and heteronuclei, as well as H-1-N-15 dipolar and H-1(N) CSA tensors are readily obtained from the high sensitivity and resolution proton-detected 3D experiments. The approach demonstrated here is expected to enable the determination of atomic-resolution structures of large protein assemblies, inaccessible by current methodologies.
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