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Comparison of 15N- and 13C-determined parameters of mobility in melittin

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Abstract

Backbone and tryptophan side-chain mobilities in the 26-residue, cytolytic peptide melittin (MLT) were investigated by 15N and 13C NMR. Specifically, inverse-detected 15N T1 and steady-state NOE measurements were made at 30 and 51 MHz on MLT at 22 °C enriched with 15N at six amide positions and in the Trp19 side chain. Both the disordered MLT monomer (1.2 mM peptide at pH 3.6 in neat water) and α-helical MLT tetramer (4.0 mM peptide at pH 5.2 in 150 mM phosphate buffer) were examined. The relaxation data were analyzed in terms of the Lipari and Szabo model-free formalism with three parameters: τm, the correlation time for the overall rotation; S2, a site-specific order parameter which is a measure of the amplitude of the internal motion; and τe, a local, effective correlation time of the internal motion. A comparison was made of motional parameters from the 15N measurements and from 13C measurements on MLT, the latter having been made here and previously [Kemple et al. (1997) Biochemistry, 36, 1678–1688]. τm and τe values were consistent from data on the two nuclei. In the MLT monomer, S2 values for the backbone N-H and Cα-H vectors in the same residue were similar in value but in the tetramer the N-H order parameters were about 0.2 units larger than the Cα-H order parameters. The Trp side-chain N-H and C-H order parameters, and τe values were generally similar in both the monomer and tetramer. Implications of these results regarding the dynamics of MLT are examined.

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References

  • Barbato, G., Ikura, M., Kay, L.E., Pastor, R.W. and Bax, A. (1992) Biochemistry, 31, 5269–5278.

    Google Scholar 

  • Brown, L.R, Lauterwein, J. and Wüthrich, K. (1980) Biochim. Biophys. Acta, 622, 231–244.

    Google Scholar 

  • Buckley, P., Edison, A.S., Kemple, M.D. and Prendergast, F.G. (1993) J. Biomol. NMR, 3, 639–652.

    Google Scholar 

  • Chandrasekhar, I., Clore, G.M., Szabo, A., Gronenborn, A.M. and Brooks, B.R. (1992) J. Mol. Biol., 226, 239–250.

    Google Scholar 

  • Clore, G.M., Szabo, A., Bax, A., Kay, L.E., Driscoll, P.C. and Gronenborn, A.M. (1990) J. Am. Chem. Soc., 112, 4989–4991.

    Google Scholar 

  • Cross, T.A. and Opella, S.J. (1983) J. Am. Chem. Soc., 105, 306–308.

    Google Scholar 

  • Daggett, V., Kollman, P.A. and Kuntz, I.D. (1991) Biopolymers, 31, 1115–1134.

    Google Scholar 

  • Dempsey, C.E. (1990) Biochim. Biophys. Acta, 1031, 143–161.

    Google Scholar 

  • Eriksson, M.A.L., Berglund, H., Härd, T. and Nilsson, L. (1993) Proteins Struct. Funct. Genet., 17, 375–390.

    Google Scholar 

  • Fadel, A.R., Jin, D.Q., Montelione, G.T. and Levy, R.M. (1995) J. Biomol. NMR, 6, 221–226.

    Google Scholar 

  • Farrow, N.A., Zhang, O., Szabo, A., Torchia, D.A. and Kay, L.E. (1995) J. Biomol. NMR, 6, 153–162.

    Google Scholar 

  • Fox, T. and Kollman, P.A. (1996) Proteins Struct. Funct. Genet., 25, 315–334.

    Google Scholar 

  • Fushman, D., Cahill, S. and Cowburn, D. (1997) J. Mol. Biol., 266, 173–194.

    Google Scholar 

  • Grzesiek, R. and Bax, A. (1993) J. Am. Chem. Soc., 115, 12593–12594.

    Google Scholar 

  • Hiyama, Y., Niu, C., Silverton, J.V., Bavoso, A. and Torchia, D.A. (1988) J. Am. Chem. Soc., 110, 2378–2383.

    Google Scholar 

  • Ishima, R., Yamasaki, K. and Nagayama, K. (1995a) J. Biomol. NMR, 6, 423–426.

    Google Scholar 

  • Ishima, R., Yamasaki, K., Saito, M. and Nagayama, K. (1995b) J. Biomol. NMR, 6, 217–220.

    Google Scholar 

  • Kay, L.E., Torchia, D.A. and Bax, A. (1989) Biochemistry, 28, 8972–8979.

    Google Scholar 

  • Kemple, M.D., Yuan, P., Nollet, K.E., Fuchs, J.A., Silva, N. and Prendergast, F.G. (1994) Biophys. J., 66, 2111–2126.

    Google Scholar 

  • Kemple, M.D., Buckley, P., Yuan, P. and Prendergast, F.G. (1997) Biochemistry, 36, 1678–1688.

    Google Scholar 

  • Kinosita, K.S., Kawato, S. and Ikegami, A. (1977) Biophys. J., 20, 289–305.

    Google Scholar 

  • Kördel, J., Skelton, N.J., Akke, M., Palmer, A.G. and Chazin, W.J. (1992) Biochemistry, 31, 4856–4866.

    Google Scholar 

  • Kördel, J. and Teleman, O. (1992) J. Am. Chem. Soc., 114, 4934–4936.

    Google Scholar 

  • Lauterwein, J., Brown, L.R. and Wüthrich, K. (1980) Biochim. Biophys. Acta, 622, 219–230.

    Google Scholar 

  • Lee, L.K., Rance, M., Chazin, W.J. and Palmer, A.G. (1997) J. Biomol. NMR, 9, 287–298.

    Google Scholar 

  • Lefèvre, J.-F., Dayie, K.T., Peng, J.W. and Wagner, G. (1996) Biochemistry, 35, 2674–2686.

    Google Scholar 

  • LeMaster, D.M. and Kushlan, D.M. (1996) J. Am. Chem. Soc., 118, 9255–9264.

    Google Scholar 

  • Lipari, G. and Szabo, A. (1982) J. Am. Chem. Soc., 104, 4546–4559.

    Google Scholar 

  • London, R.E. and Avitabile, J. (1978) J. Am. Chem. Soc., 100, 7159–7165.

    Google Scholar 

  • Mandel, A.M., Akke, M. and Palmer, A.G. (1995) J. Mol. Biol., 246, 144–163.

    Google Scholar 

  • Palmer, A.G. and Case, D.A. (1992) J. Am. Chem. Soc., 114, 9059–9067.

    Google Scholar 

  • Palmer, A.G., Hochstrasser, R.A., Millar, D.P., Rance, M. and Wright, P.E. (1993) J. Am. Chem. Soc., 115, 6333–6345.

    Google Scholar 

  • Peng, J.W. and Wagner, G. (1995) Biochemistry, 34, 16733–16752.

    Google Scholar 

  • Schneider, D.M., Dellow, M.J. and Wand, A.J. (1992) Biochemistry, 31, 3645–3652.

    Google Scholar 

  • Shaka, A.J., Keeler, J., Frenkiel, T. and Freeman, R. (1983) J. Magn. Reson., 52, 335–338.

    Google Scholar 

  • Shaka, A.J., Barker, P.B. and Freeman, R. (1985) J. Magn. Reson., 64, 547–552.

    Google Scholar 

  • Smith, L.J., Mark, A.E., Dobson, C.M. and van Gunsteren, W.F. (1995a) Biochemistry, 34, 10918–10931.

    Google Scholar 

  • Smith, P.E., Schaik, R.C., Szyperski, T., Wüthrich, K. and van Gunsteren, W.F. (1995b) J. Mol. Biol., 246, 356–365.

    Google Scholar 

  • Stone, M.J., Fairbrother, W.J., Palmer, A.G., Reizer, J., Saier, M.H. and Wright, P.E. (1992) Biochemistry, 31, 4394–4406.

    Google Scholar 

  • Stone, M.J., Chandrasekhar, K., Holmgren, A., Wright, P.E. and Dyson, H.J. (1993) Biochemistry, 32, 426–435.

    Google Scholar 

  • Terwilliger, T.C. and Eisenberg, D. (1982a) J. Biol. Chem., 257, 6010–6015.

    Google Scholar 

  • Terwilliger, T.C. and Eisenberg, D. (1982b) J. Biol. Chem., 257, 6016–6022.

    Google Scholar 

  • Torchia, D.A., Nicholson, L.K., Cole, H.B.R. and Kay, L.E. (1993) In NMR of Proteins (Eds., Clore, G.M. and Gronenborn, A.M.), CRC Press, Boca Raton, FL, pp. 190–219.

    Google Scholar 

  • Wand, A.J., Urbauer, J.L., McEvoy, R.P. and Bieber, R.J. (1996) Biochemistry, 35, 6116–6125.

    Google Scholar 

  • Weaver, A.J., Kemple, M.D. and Prendergast, F.G. (1989) Biochemistry, 28, 8624–8639.

    Google Scholar 

  • Wishart, D.S., Sykes, B.D. and Richards, F.M. (1991) J. Mol. Biol., 222, 311–333.

    Google Scholar 

  • Wishart, D.S., Bigam, C.G., Yao, J., Abildgaard, F., Dyson, H.J., Oldfield, E., Markley, J.L. and Sykes, B.D. (1995) J. Biomol. NMR, 6, 135–140.

    Google Scholar 

  • Yamasaki, K., Saito, M., Oobatake, M. and Kanaya, S. (1995) Biochemistry, 34, 6587–6601.

    Google Scholar 

  • Yamazaki, T., Muhandiram, R. and Kay, L.E. (1994) J. Am. Chem. Soc., 116, 8266–8278.

    Google Scholar 

  • Yuan, P., Fisher, P.J., Prendergast, F.G. and Kemple, M.D. (1996) Biophys. J., 70, 2223–2238.

    Google Scholar 

  • Zheng, Z., Czaplicki, J. and Jardetzky, O. (1995) Biochemistry, 34, 5212–5223.

    Google Scholar 

  • Zhu, L., Kemple, M.D., Yuan, P. and Prendergast, F.G. (1995) Biochemistry, 34, 13196–13202.

    Google Scholar 

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Authors and Affiliations

  1. Department of Physics, Indiana University Purdue, University Indianapolis, 402 North Blackford Street, Indianapolis, IN, 46202-3273, U.S.A

    Lingyang Zhu & Marvin D. Kemple

  2. Department of Pharmacology, Mayo Foundation, Rochester, MN, 55905, U.S.A

    Franklyn G. Prendergast

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  1. Lingyang Zhu
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  2. Franklyn G. Prendergast
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  3. Marvin D. Kemple
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Zhu, L., Prendergast, F.G. & Kemple, M.D. Comparison of 15N- and 13C-determined parameters of mobility in melittin. J Biomol NMR 12, 135–144 (1998). https://doi.org/10.1023/A:1008210613711

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