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The use of heteronuclear cross-polarization for backbone assignment of 2H-, 15N- and 13C-labeled proteins: A pulse scheme for triple-resonance 4D correlation of sequential amide protons and 15N

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Summary

A new four-dimensional pulse scheme is described for the main-chain assignment of proteins by means of the J connectivity of the amide proton and nitrogen resonances of adjacent residues. Since the new experiment, 4D CP-HN(COCA)NH, involves heteronuclear cross-polarization for magnetization transfer from 13C=O to 15N via 13Cα, a relatively strong WALTZ-16 decoupling rf field is applied to 13Cα during magnetization transfer. Consequently, 13Cα is effectively decoupled from its attached 2H in the case of deuterated proteins, in the absence of a decoupling rf field for 2H. This efficiently improves the sensitivity of the experiment through 13C line narrowing. The experiment was performed on a randomly 60% deuterated protein, and the sensitivity of the final 4D spectrum was found to be excellent.

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References

  • Abragam, A. (1961) The Principles of Nuclear Magnetism, Clarendon Press, Oxford.

    Google Scholar 

  • Bax, A. and Grzesiek, S. (1993) Acc. Chem. Res. 26, 131–138.

    Google Scholar 

  • Boucher, W., Raine, A.R.C. and Laue, E.D. (1991) In Computational Aspects of the Study of Biological Macromolecules by Nuclear Magnetic Resonance Spectroscopy (Eds, Hoch, J.C., Poulsen, F.M. and Redfield, C.) NATO ASI Series, Vol. 225, Plenum Press, New York, NY, p. 87.

    Google Scholar 

  • Ernst, M., Griesinger, C. and Ernst, R.R. (1991) Mol. Phys., 74, 219–252.

    Google Scholar 

  • Grzesiek, S., Anglister, J., Ren, H. and Bax, A. (1993) J. Am. Chem. Soc., 115, 4369–4370.

    Google Scholar 

  • Ikura, M., Kay, L.E. and Bax, A. (1990) Biochemistry, 29, 4659–4667.

    Google Scholar 

  • Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.

    Google Scholar 

  • Kushlan, D.M. and LeMaster, D.M. (1993) J. Biomol. NMR, 3, 701–708.

    Google Scholar 

  • Laue, E.D., Mayger, M.R., Skilling, J. and Staunton, J. (1986) J. Magn. Reson., 68, 14–29.

    Google Scholar 

  • Majumdar, A., Wang, H., Morshauser, R.C. and Zuiderweg, E.R.P. (1993) J. Biomol. NMR, 3, 387–397.

    Google Scholar 

  • Marion, D., Ikura, M. and Bax, A. (1989) J. Magn. Reson. 84, 425–430.

    Google Scholar 

  • Piotto, M., Saudek, V. and Sklenář, V., J. Biomol. NMR, 2, 661–665.

  • Richardson, J.M., Clowes, R.T., Boucher, W., Domaille, P.J., Hardman, C.H., Keeler, J. and Laue, E.D. (1993) J. Magn. Reson. Ser. B, 101, 223–227.

    Google Scholar 

  • Schleucher, J., Schwendinger, M., Sattler, M., Schmidt, P., Glaser, S.J., Sørensen, O.W. and Griesinger, C. (1994) J. Biomol. NMR, 4, 301–306.

    Google Scholar 

  • Vincent, S.J.F., Zwahlen, C. and Bodenhausen, G. (1993) J. Am. Chem. Soc., 115, 9202–9209.

    Google Scholar 

  • Yamazaki, T., Lee, W., Revington, M., Mattiello, D., Dahlquist, F.W., Arrowsmith, C.H. and Kay, L.E. (1994) J. Am. Chem. Soc., 116, 6464–6465.

    Google Scholar 

  • Zuiderweg, E.R.P. (1990) J. Magn. Reson., 89, 533–542.

    Google Scholar 

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Authors and Affiliations

  1. Institute for Protein Research, Osaka University, Suita, Osaka 565, Japan

    Masahiro Shirakawa, Masato Shimizu & Yoshimasa Kyogoku

  2. Bruker Japan, 21-5, Ninomiya 3-chome, Tsukuba, Ibaraki 305, Japan

    Markus Wälchli

Authors
  1. Masahiro Shirakawa
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  2. Markus Wälchli
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  3. Masato Shimizu
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  4. Yoshimasa Kyogoku
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Shirakawa, M., Wälchli, M., Shimizu, M. et al. The use of heteronuclear cross-polarization for backbone assignment of 2H-, 15N- and 13C-labeled proteins: A pulse scheme for triple-resonance 4D correlation of sequential amide protons and 15N. J Biomol NMR 5, 323–326 (1995). https://doi.org/10.1007/BF00211761

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  • DOI: https://doi.org/10.1007/BF00211761

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