Resolution and measurement of heteronuclear dipolar couplings of a noncrystalline protein immobilized in a biological supramolecular assembly by proton-detected MAS solid-state NMR spectroscopy.
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.
['Park|Sang Ho|SH|', 'Yang|Chen|C|', 'Opella|Stanley J|SJ|', 'Mueller|Leonard J|LJ|']
D000465:Algorithms D025561:Bacteriophage Pf1 D036022:Capsid Proteins D004279:DNA, Viral D003903:Deuterium D005998:Glycine D009682:Magnetic Resonance Spectroscopy D009587:Nitrogen Isotopes D019906:Nuclear Magnetic Resonance, Biomolecular D011506:Proteins D011522:Protons
Perdeuteration Fast MAS Pf1 bacteriophage Proton detection Separated local field spectroscopy Three-dimensional NMR Two-dimensional NMR Variable contact time cross-polarization