Summary
The solution structure of gurmarin was studied by two-dimensional proton NMR spectroscopy at 600 MHz. Gurmarin, a 35-amino acid residue polypeptide recently discovered in an Indian-originated tree Gymnema sylvestre, selectively suppresses the neural responses of rat to sweet taste stimuli. Sequence-specific protons. The three-dimensional solution structure was determined by simulated-annealing calculations on the basis of 135 interproton distance constraints derived from NOEs, six distance constraints for three hydrogen bonds and 16 dihedral angle constraints derived from coupling constants. A total of 10 structures folded into a well-defined structure with a triple-stranded antiparallel β-sheet. The average rmsd values between any two structures were 1.65±0.39 Å for the backbone atoms (N, Cα, C) and 2.95±0.27 Å for all heavy atoms. The positions of the three disulfide bridges, which could not be deterermined chemically, were estimated to be Cys3–Cys18, Cys10–Cys23 and Cys17–Cys33 on the basis of the NMR distance constraints. This disulfide bridge pattern in gurmarin turned out to be analogous to that in ω-conotoxin and Momordica charantia trypsin inhibitor-II, and the topology of folding was the same as that in ω-conotoxin.
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Abbreviations
- DQF-COSY:
-
double-quantum-filtered correlated spectroscopy
- HOHAHA:
-
homonuclear Hartmann-Hahn spectroscopy
- NOESY:
-
nuclear Overhauser enhancement spectroscopy
- ppm:
-
parts per million; rmsd, root-mean-square deviation
- TSP:
-
3-(trimethylsilyl)-2,2,3,3-tetradeutero-propionate
References
Bax, A. and Davis, D.G. (1985) J. Magn. Reson. 65, 355–360.
Edgeworth, P. (1847–48) Proc. Linnean Soc. London, 7, 351–352.
Glaser, D., Hellekant, G., Brouwer, J.N. and Van derWel, H. (1984) Chem. Senses, 8, 367–374.
Hara, S., Makino, J. and Ikenaka, T. (1989) J. Biochem., 105, 88–92.
Hellekant, G. (1976) Chem. Senses Flavour, 2, 85–95.
Hellekant, G. and Gopal, V. (1976) Acta Physiol. Scand., 98, 136–142.
Hooper, D. (1887) Nature, 35, 565–567.
Imoto, T., Miyasaka, A., Ishima, R. and Akasaka, K. (1991) Comput. Biochem. Physiol. A, 100, 309–314.
Jeener, T., Meier, B.H., Bachmann, P. and Ernst, R.R. (1979) J. Chem. Phys. 71, 4546–4553.
Kamei, K., Takano, R., Miyasaka, A., Imoto, T. and Hara, S. (1992) J. Biochem., 111, 109–112.
Macura, C., Huang, Y., Suter, D. and Ernst, R.R. (1981) J. Magn. Reson., 43, 259–281.
McCleskey, E.W., Fox, A.P., Feldman, D.H., Cruz, L.J., Olivera, B.M., Tsien, R.W. and Yoshikami, D. (1987) Proc. Natl. Acad. Sci. USA, 84, 4327–4331.
Nakai, T., Kidera, A. and Nakamura, H. (1993) J. Biomol. NMR, 3, 19–40.
Nilges, M., Clore, G.M. and Gronenborn, A.M. (1988) FEBS Lett., 229, 317–324.
Nishiuchi, Y., Kumagaye, K., Noda, Y., Watanabe, T. and Sakakibara, S. (1986) Biopolymers, 25, S61–68.
Pallaghy, P.K., Duggan, B.M., Pennington, M.W. and Norton, R.S. (1993) J. Mol. Biol., 234, 405–420.
Rance, M., Sørensen, O.W., Bodenhausen, G., Wagner, G., Ernst, R.R. and Wüthrich, K. (1983) Biochem. Biophys. Res. Commun., 117, 479–485.
Wagner, G., Braun, W., Havel, T.F., Schaumann, T., Gō, N. and Wüthrich, K. (1987) J. Mol. Biol., 196, 611–639.
Weiner, S.J., Kollman, P.A., Nguyen, D.T. and Case, D.A. (1986) J. Comput. Chem., 7, 230–252.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY.
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Arai, K., Ishima, R., Morikawa, S. et al. Three-dimensional structure of gurmarin, a sweet taste-suppressing polypeptide. J Biomol NMR 5, 297–305 (1995). https://doi.org/10.1007/BF00211756
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DOI: https://doi.org/10.1007/BF00211756