Arginine side chain assignments in uniformly 15N-labeled proteins using the novel 2D HE(NE)HGHH experiment |
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Authors: | Maurizio Pellecchia Gerhard Wider Hideo Iwai Kurt Wüthrich |
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Affiliation: | (1) Eidgenössische Technische Hochschule-Hönggerberg, Institut für Molekularbiologie und Biophysik, CH-8093 Zürich, Switzerland |
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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 between1H, 1Hand 1H are established on the basis of3J(15N,1H) heteronuclear scalarcoupling constants, and sequence-specific assignments are obtained by overlapof these fragments with 1H 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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Keywords: | NMR assignments of arginine Protein structure determination Protein– DNA recognition |
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