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Structure of the cyclic peptide [W8S]contryphan Vn: effect of the tryptophan/serine substitution on trans–cis proline isomerization

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Abstract

The structural characterization of [W8S]contryphan Vn, an analogue of Contryphan Vn with tryptophan 8 substituted with a serine residue (W8S), was performed by NMR spectroscopy, molecular dynamics simulations and fluorescence spectroscopy. Contryphan Vn, a bioactive cyclic peptide from the venom of the cone snail Conus ventricosus, contains an S–S bridge between two cysteines and a d-tryptophan. Like other Contryphans, [W8S]contryphan Vn has proline 7 isomerized trans, while the proline 4 has nearly equivalent populations of cis and trans configurations. The thermodynamic and kinetic parameters of the trans–cis isomerization of proline 4 were measured. The isomers of [W8S]contryphan Vn with proline 4 in cis and trans show structural differences. The absence of the salt bridge between the same Asp2 and Lys6, present in Contryphan Vn, may be attributed to the lack of the hydrophobic side chain of Trp8 where it likely protects the electrostatic interactions. These results may contribute to identifying, in these cyclic peptides, the structural determinants of the mechanism of proline trans–cis isomerization, this being also an important step in protein folding.

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Abbreviations

TFA:

Trifluoroacetic acid

HMQC:

Heteronuclear multiple quantum coherence

HSQC:

Heteronuclear single quantum coherence

ROESY:

Rotating-frame Overhauser spectroscopy

TOCSY:

Total correlation spectroscopy

DOSY:

Diffusion-ordered spectroscopy

TFE:

Trifluoroethanol

PME:

Particle-mesh Ewald method

TSP:

Trimethylsilyl propionic acid

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Acknowledgments

The technical assistance of Mr. Fabio Bertocchi is gratefully acknowledged.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Chemical Sciences and Technologies, University of Rome “Tor Vergata”, Via della Ricerca Scientifica 1, 00133, Rome, Italy

    Ridvan Nepravishta, Walter Mandaliti, Sonia Melino, Tommaso Eliseo & Maurizio Paci

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  1. Ridvan Nepravishta
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  2. Walter Mandaliti
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  3. Sonia Melino
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  4. Tommaso Eliseo
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  5. Maurizio Paci
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Corresponding author

Correspondence to Maurizio Paci.

Additional information

R. Nepravishta and W. Mandaliti have equally contributed to this work.

Electronic supplementary material

Supplementary Table 1. Assignment of the NMR resonances of [W8S]contryphan Vn spectra. The chemical shift values of 1H, 15N, and 13C resonances are reported. Labels A and B indicate the resonances due to the isomers of Proline 4 cis and trans, respectively.

Supplementary Table 2a and b. NMR Structural Statistics from XPLOR. Family of 20 Structures for cis-[W8S]contryphan Vn a) and family of 20 Structures trans-[W8S]contryphan Vn b).

Supplementary Fig. 1. Heteronuclear 1H-13C HSQC NMR spectrum of [W8S]contryphan Vn. In figure, some of 13C resonances assigned are shown. The complete list of assignments is reported in Supplementary Table 1.

Supplementary Fig. 2. DOSY NMR of [W8S]contryphan Vn in H2O/D2O 90/10 (v/v) at pH 3.0.

Supplementary Fig. 3a. Comparison between phi angles of cis-[W8S]contryphan Vn (black) and trans-[W8S]contryphan Vn (gray).

Supplementary Fig. 3b. Comparison between psi angles of cis-[W8S]contryphan Vn (black) and trans-[W8S]contryphan Vn (gray).

Supplementary Fig. 3c. Comparison between phi angles of Contryphan Vn (black) and cis-[W8S]contryphan Vn (gray).

Supplementary Fig. 3d. Comparison between psi angles of Contryphan Vn (black) and cis-[W8S]contryphan Vn (gray).

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 132 kb)

Supplementary material 2 (PDF 128 kb)

Supplementary material 3 (PDF 104 kb)

Supplementary material 4 (PDF 101 kb)

Supplementary material 5 (PDF 102 kb)

Supplementary material 6 (PDF 101 kb)

Supplementary material 7 (PDF 153 kb)

Supplementary material 8 (PDF 84 kb)

Supplementary material 9 (PDF 84 kb)

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Nepravishta, R., Mandaliti, W., Melino, S. et al. Structure of the cyclic peptide [W8S]contryphan Vn: effect of the tryptophan/serine substitution on trans–cis proline isomerization. Amino Acids 46, 2841–2853 (2014). https://doi.org/10.1007/s00726-014-1841-9

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