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Application of multiple-quantum line narrowing with simultaneous 1H and 13C constant-time scalar-coupling evolution in PFG-HACANH and PFG-HACA(CO)NH triple-resonance experiments

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Abstract

Many triple-resonance experiments make use of one-bond heteronuclear scalar couplings toestablish connectivities among backbone and/or side-chain nuclei. In medium-sized(15–30 kDa) proteins, short transverse relaxation times of Cα single-quantum stateslimit signal-to-noise (S/N) ratios. These relaxation properties can be improved usingheteronuclear multiple-quantum coherences (HMQCs) instead of heteronuclear single-quantumcoherences (HSQCs) in the pulse sequence design. In slowly tumbling macromolecules, theseHMQCs can exhibit significantly better transverse relaxation properties than HSQCs.However, HMQC-type experiments also exhibit resonance splittings due to multiple two- andthree-bond homo- and heteronuclear scalar couplings. We describe here a family of pulsed-field gradient (PFG) HMQC-type triple-resonance experiments using simultaneous 1H and13C constant-time (CT) periods to eliminate the t1 dependence of these scalar couplingeffects. These simultaneous CT PFG-(HA)CANH and PFG-(HA)CA(CO)NH HMQC-typeexperiments exhibit sharper resonance line widths and often have better S/N ratios than thecorresponding HSQC-type experiments. Results on proteins ranging in size from 6 to 30 kDashow average methine CαH HMQC:HSQC enhancement factors of 1.10 ± 0.15, withabout 40% of the cross peaks exhibiting better S/N ratios in the simultaneous CT-HMQCversions compared with the HSQC versions.

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References

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

    Google Scholar 

  • Billeter, M., Neri, D., Otting, G., Qian, Y.Q. and Wüthrich, K. (1992) J. Biomol. NMR, 2, 257–274.

    Google Scholar 

  • Boucher, W., Laue, E.D., Campbell-Burk, S. and Domaille, P.J. (1992a) J. Am. Chem. Soc., 114, 2262–2264.

    Google Scholar 

  • Boucher, W., Laue, E.D., Campbell-Burk, S.L. and Domaille, P.J. (1992b) J. Biomol. NMR, 2, 631–637.

    Google Scholar 

  • Clowes, R.T., Boucher, W., Hardman, C.H., Domaille, P.J. and Laue, E.D. (1993) J. Biomol. NMR, 3, 349–354.

    Google Scholar 

  • Clubb, R.T. and Wagner, G. (1992) J. Biomol. NMR, 2, 389–394.

    Google Scholar 

  • Clubb, R.T., Thanabal, V. and Wagner, G. (1992a) J. Magn. Reson., {vn97}, 213–217.

    Google Scholar 

  • Clubb, R.T., Thanabal, V. and Wagner, G. (1992b) J. Biomol. NMR, {vn2}, 203–210.

    Google Scholar 

  • Ernst, R.R., Bodenhausen, G. and Wokaun, A. (1987) Principles of NMR in One and Two Dimensions, Clarendon Press, Oxford, U.K.

    Google Scholar 

  • Farmer II, B.T. and Venters, R.A. (1995) J. Am. Chem. Soc., 117, 4187–4188.

    Google Scholar 

  • Feng, W., Rios, C.B. and Montelione, G.T. (1996) J. Biomol. NMR, {vn8}, 98–104.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1992a) J. Am. Chem. Soc., 114, 6291–6293.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1992b) J. Magn. Reson., 99, 201–207.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1993) J. Biomol. NMR, 3, 185–204.

    Google Scholar 

  • Grzesiek, S., Anglister, J. and Bax, A. (1993a) J. Magn. Reson., B101, 114–119.

    Google Scholar 

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

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1995) J. Biomol. NMR, 6, 335–339.

    Google Scholar 

  • Grzesiek, S., Kuboniwa, H., Hinck, A.P. and Bax, A. (1995) J. Am. Chem. Soc., 117, 5312–5315.

    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 

  • Kay, L.E., Ikura, M. and Bax, A. (1991) J. Magn. Reson., 91, 84–92.

    Google Scholar 

  • Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663–10665.

    Google Scholar 

  • Kuboniwa, H., Grzesiek, S., Delaglio, F. and Bax, A. (1994) J. Biomol. NMR, 4, 871–878.

    Google Scholar 

  • Logan, T.M., Olejniczak, E.T., Xu, R.X. and Fesik, S.W. (1992) FEBS Lett., 314, 413–418.

    Google Scholar 

  • Lyons, B.A. and Montelione, G.T. (1993) J. Magn. Reson., B101, 206–209.

    Google Scholar 

  • Lyons, B.A., Tashiro, M., Cedergren, L., Nilsson, B. and Montelione, G.T. (1993) Biochemistry, 32, 7839–7845.

    Google Scholar 

  • Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 84, 393–399.

    Google Scholar 

  • Montelione, G.T. and Wagner, G. (1989) J. Am. Chem. Soc., 111, 5474–5475.

    Google Scholar 

  • Montelione, G.T. and Wagner, G. (1990) J. Magn. Reson., 87, 183–188.

    Google Scholar 

  • Montelione, G.T., Lyons, B.A., Emerson, S.D. and Tashiro, M. (1992) J. Am. Chem. Soc., 114, 10974–10975.

    Google Scholar 

  • Norwood, T.J. (1992) Prog. NMR Spectrosc., 24, 295–375.

    Google Scholar 

  • Olejniczak, E.T., Xu, R.X., Petros, A.M. and Fesik, S.W. (1992) J. Magn. Reson., 100, 444–450.

    Google Scholar 

  • Qian, X.-Y., Chien, C.-Y., Lu, Y., Montelione, G.T. and Krug, R.M. (1995) RNA, 1, 948–956.

    Google Scholar 

  • Seip, S., Balbach, J. and Kessler, H. (1992) J. Magn. Reson., 100, 406–410.

    Google Scholar 

  • Shaka, A.J., Barker, P.B. and Freeman, R. (1985) J. Magn. Reson., {vn64}, 547–552.

    Google Scholar 

  • Wittekind, M. and Mueller, L. (1993) J. Magn. Reson., B101, 201–205.

    Google Scholar 

  • Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY, U.S.A.

    Google Scholar 

  • Yamazaki, T., Lee, W., Revington, M., Mattiello, D.L., Dahlquist, F.W., Arrowsmith, C.H. and Kay, L.E. (1994) J. Am. Chem. Soc., {vn116}, 6464–6465.

    Google Scholar 

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Authors and Affiliations

  1. Center for Advanced Biotechnology and Medicine and Department of Molecular Biology and Biochemistry, Rutgers University, 679 Hoes Lane, Piscataway, NJ, 08854-5638, U.S.A

    G.V.T. Swapna, Carlos B. Rios, Zhigang Shang & Gaetano T. Montelione

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  1. G.V.T. Swapna
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  2. Carlos B. Rios
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  3. Zhigang Shang
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  4. Gaetano T. Montelione
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Swapna, G., Rios, C.B., Shang, Z. et al. Application of multiple-quantum line narrowing with simultaneous 1H and 13C constant-time scalar-coupling evolution in PFG-HACANH and PFG-HACA(CO)NH triple-resonance experiments. J Biomol NMR 9, 105–111 (1997). https://doi.org/10.1023/A:1018683920602

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