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Methods for sequential resonance assignment in solid, uniformly 13C, 15N labelled peptides: Quantification and application to antamanide

Detken, Andreas ; Hardy, Edme ; Ernst, Matthias ; Kainosho, Masatsune ; Kawakami, Toru ; Aimoto, Saburo ; Meier, Beat

In: Journal of Biomolecular NMR, 2001, vol. 20, no. 3, p. 203-221

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    Title
    • Methods for sequential resonance assignment in solid, uniformly 13C, 15N labelled peptides: Quantification and application to antamanide
    Author
    • Detken, Andreas. Laboratory of Physical Chemistry, ETH Hönggerberg, CH-8093, Zürich, Switzerland
    • Hardy, Edme. Laboratory of Physical Chemistry, ETH Hönggerberg, CH-8093, Zürich, Switzerland
    • Ernst, Matthias. Laboratory of Physical Chemistry, ETH Hönggerberg, CH-8093, Zürich, Switzerland
    • Kainosho, Masatsune. CREST, Japan Science and Technology Corporation, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Tokyo, Japan
    • Kawakami, Toru. Institute for Protein Research, Osaka University, 3-2 Yamadaoka, 565-0871, Suita, Osaka, Japan
    • Aimoto, Saburo. Institute for Protein Research, Osaka University, 3-2 Yamadaoka, 565-0871, Suita, Osaka, Japan
    • Meier, Beat. Laboratory of Physical Chemistry, ETH Hönggerberg, CH-8093, Zürich, Switzerland
    Document Type
    • Postprint
    Language
    • English
    Published in
    • Journal of Biomolecular NMR, 2001, vol. 20, no. 3, p. 203-221. Kluwer Academic Publishers
    Other Version
    OAI-PMH Identifier
    • oai:doc.rero.ch:317002
    Summary
    The application of adiabatic polarization-transfer experiments to resonance assignment in solid, uniformly 13C-15N-labelled polypeptides is demonstrated for the cyclic decapeptide antamanide. A homonuclear correlation experiment employing the DREAM sequence for adiabatic dipolar transfer yields a complete assignment of the Cα and aliphatic side-chain 13C resonances to amino acid types. The same information can be obtained from a TOBSY experiment using the recently introduced P91 12 TOBSY sequence, which employs the J couplings as a transfer mechanism. A comparison of the two methods is presented. Except for some aromatic phenylalanine resonances, a complete sequence-specific assignment of the 13C and 15N resonances in antamanide is achieved by a series of selective or broadband adiabatic triple-resonance experiments. Heteronuclear transfer by adiabatic-passage Hartmann-Hahn cross polarization is combined with adiabatic homonuclear transfer by the DREAM and rotational-resonance tickling sequences into two- and three-dimensional experiments. The performance of these experiments is evaluated quantitatively