Abstract
Two-dimensional (15)N chemical shift/(1)H chemical shift and three-dimensional (1)H-(15)N dipolar coupling/(15)N chemical shift/(1)H chemical shift MAS solid-state NMR correlation spectra of the filamentous bacteriophage Pf1 major coat protein show single-site resolution in noncrystalline, intact-phage preparations. The high sensitivity and resolution result from (1)H detection at 600MHz under 50kHz magic angle spinning using ∼0.5mg of perdeuterated and uniformly (15)N-labeled protein in which the exchangeable amide sites are partially or completely back-exchanged (reprotonated). Notably, the heteronuclear (1)H-(15)N dipolar coupling frequency dimension is shown to select among (15)N resonances, which will be useful in structural studies of larger proteins where the resonances exhibit a high degree of overlap in multidimensional chemical shift correlation spectra.
- algorithms,bacteriophage
- bacteriophage,proteins,proton
- contact
- cross-polarization,viral
- detection,protons,separated
- field
- isotopes,nuclear
- local
- magnetic
- mas,glycine,magnetic
- nmr,two-dimensional
- nmr,variable
- pf1,biomolecular,capsid
- proteins,dna,deuterium,fast
- resonance
- resonance,perdeuteration,pf1
- spectroscopy,nitrogen
- spectroscopy,three-dimensional
- time
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