Assignment of amide proton signals by combined evaluation of HN, NN and HNCA MAS-NMR correlation spectra.
In this paper, we present a strategy for the (1)H(N) resonance assignment in solid-state magic-angle spinning (MAS) NMR, using the alpha-spectrin SH3 domain as an example. A novel 3D triple resonance experiment is presented that yields intraresidue H(N)-N-C(alpha) correlations, which was essential for the proton assignment. For the observable residues, 52 out of the 54 amide proton resonances were assigned from 2D ((1)H-(15)N) and 3D ((1)H-(15)N-(13)C) heteronuclear correlation spectra. It is demonstrated that proton-driven spin diffusion (PDSD) experiments recorded with long mixing times (4 s) are helpful for confirming the assignment of the protein backbone (15)N resonances and as an aid in the amide proton assignment.
van Rossum
Barth-Jan
BJ
Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle Str. 10, D-13125, Berlin, Germany.
Castellani
Federica
F
Pauli
Jutta
J
Rehbein
Kristina
K
Hollander
J
J
de Groot
Huub J M
HJ
Oschkinat
Hartmut
H
D000577:Amides / Q000737:chemistry*; D002247:Carbon Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008957:Models, Genetic; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011522:Protons; D013049:Spectrin / Q000737:chemistry; D013696:Temperature; D018909:src Homology Domains
D000577:Amides; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011522:Protons; D013049:Spectrin