Assignment of amide proton signals by combined evaluation of HN,NN and HNCA MAS-NMR correlation spectra |
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Authors: | van Rossum Barth-Jan Castellani Federica Pauli Jutta Rehbein Kristina Hollander J de Groot Huub J M Oschkinat Hartmut |
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Affiliation: | (1) Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle Str. 10, D-13125 Berlin, Germany;(2) Present address: BAM, Richard-Willstätter-Str. 11, D-12489 Berlin, Germany |
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Abstract: | In this paper, we present a strategy for the 1HN resonance assignment in solid-state magic-angle spinning (MAS) NMR, using the -spectrin SH3 domain as an example. A novel 3D triple resonance experiment is presented that yields intraresidue HN-N-C correlations, which was essential for the proton assignment. For the observable residues, 52 out of the 54 amide proton resonances were assigned from 2D (1H-15N) and 3D (1H-15N-13C) 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 15N resonances and as an aid in the amide proton assignment. |
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Keywords: | assignment dipolar correlation spectroscopy magic-angle-spinning SH3 domain solid-state MAS NMR |
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