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Arginine side chain assignments in uniformly 15N-labeled proteins using the novel 2D HE(NE)HGHH experiment

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Abstract

A novel 2D NMR experiment, 2D HE(NE)HGHH, is presented for the assignment ofarginine side chain 1H and 15N resonances inuniformly 15N-labeled proteins. Correlations between1Hε, 1Hγand 1Hη are established on the basis of3J(15N,1H) heteronuclear scalarcoupling constants, and sequence-specific assignments are obtained by overlapof these fragments with 1Hγ chemical shiftsobtained by assignment procedures starting from the polypeptide backbone.Since guanidino protons exchange quite rapidly with the bulk water, the 2DHE(NE)HGHH pulse scheme has been optimized to avoid saturation and dephasingof the water magnetization during the course of the experiment. As anillustration, arginine side chain assignments are presented for two uniformly15N-labeled proteins of 7 and 23 kDa molecular weight.

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Authors and Affiliations

  1. Eidgenössische Technische Hochschule-Hönggerberg, Institut für Molekularbiologie und Biophysik, CH-8093, Zürich, Switzerland

    Maurizio Pellecchia, Gerhard Wider, Hideo Iwai & Kurt Wüthrich

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  1. Maurizio Pellecchia
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  2. Gerhard Wider
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  3. Hideo Iwai
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  4. Kurt Wüthrich
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Pellecchia, M., Wider, G., Iwai, H. et al. Arginine side chain assignments in uniformly 15N-labeled proteins using the novel 2D HE(NE)HGHH experiment. J Biomol NMR 10, 193–197 (1997). https://doi.org/10.1023/A:1018381109524

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