Article | Published:

Three-dimensional structure of the RGD-containing neurotoxin homologue dendroaspin

Nature Structural Biologyvolume 1pages802807 (1994) | Download Citation

Subjects

Abstract

Dendroaspin is a short chain neurotoxin homologue from the venom of Elapidae snakes, which lacks neurotoxicity. Unlike neurotoxins, it contains an Arg-Gly-Asp-(RGD)-motif and functions as an inhibitor of platelet aggregation and platelet adhesion with comparable potency to the disintegrins from the venoms of Viperidae. We have determined the structure of dendroaspin in solution using NMR spectroscopy. The structure contains a core similar to that of short chain neurotoxins, but with a novel arrangement of loops and a solvent-exposed RGD-motif. Dendroaspin is thus an integrin antagonist with a well defined fold different from that of the disintegrins, based on the neurotoxin scaffold.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Joubert, F.J. & Taljaard, N. Some properties and the complete primary structures of two reduced and S-carboxymethylated polypeptides (S5C1 and S5C10) from Dendroaspis jamesoni Kaimosae (Jameson's mamba) venom, Bioch. biophys. Acta 579, 228–233 (1979).

  2. 2

    McDowell, R.S. et at. Mambin, a potent glycoprotein IIb-IIIa antagonist and platelet aggregation inhibitor structurally related to the short neurotoxins, Biochemistry 31, 4766–4772 (1992).

  3. 3

    Williams, J.A., Lu, X., Rahman, S., Keating, C. & Kakkar, V. Dendroaspin; a potent integrin receptor inhibitor from the venoms of Dendroaspis viridis and D. jamesonii, Biochem. Soc. Trans. 21, 73S (1992).

  4. 4

    Dufton, M.J. & Hider, R.C. Conformational properties of the neurotoxins and cytotoxins isolated from elapid snake venoms, Crit. Rev. Biochem. 14, 130–136 (1983).

  5. 5

    Endo, T. & Tamiya, N. in Snake Toxins (ed. Harvey A.L.)165–222 (Pergamon, New York 1991).

  6. 6

    Dennis, M.S. et al. Platelet glycoprotein IIb-IIIa antagonists from snake venoms: evidence for a family of platelet aggregation inhibitors, Proc. natn. Acad. Sci. U.S.A. 87, 2471–2475 (1990).

  7. 7

    Gould, R.J. et al. Disintegrins, a family of integrin inhibitory proteins from viper venoms, Proc. soc. exp. biol. Med. 195, 168–171 (1990).

  8. 8

    Ruoslahti, E. & Pierschbacher, M.D. New perspectives in cell adhesion - RGD and integrins, Science 227, 491–479.(1987).

  9. 9

    Lu, X., Deadman, J.J., Williams, J.A., Kakkar, V. & Rahman, S. Synthetic RGD peptides derived from the adhesive domains of snake-venom proteins: evaluation as inhibitors of platelet aggregation, Biochem. J. 296, 21–24 (1993).

  10. 10

    Wuthrich, K. NMR of proteins and nucleic acids, (Wiley-lnterscience, New York 1986).

  11. 11

    Sutcliffe, M.J. & Dobson, C.M. Relaxation data in NMR structure determination - model calculations for the lysozyme-Gd3+ complex. Proteins 10, 117–129 (1991).

  12. 12

    Rees, B., Bilwes, A., Samana, J.P. & Moras, D., Cardiotoxin VII4 from Naja mossambica mossambica. the refined crystal structure, J. molec. Biol. 214, 281–297 (1990).

  13. 13

    Smith, J.L., Corfield, P.W. R., Hendrickson, W.A. & Low, B.W. Refinement at 1.4 Å resolution of a model of erabutoxin b-treatment of ordered solvent and discrete disorder. Acta crystallogr. A 44, 357–358 (1988).

  14. 14

    Adler, M., Lazarus, R.A., Dennis, M.S. & Wagner, G. Solution structure of kistrin, a potent platelet aggregation inhibitor and glycoprotein IIb-IIIa antagonist. Science 253, 445–448 (1991).

  15. 15

    Cooke, R.M. et al. The solution structure of echistatin: evidence for disulphide bond rearrangement in similar snake toxins. Prot. Engng 5, 473–477 (1992).

  16. 16

    Saudek, V., Atkinson, R.A. & Pelton, J.T Three dimensional structure of echistatin, the smallest active RGD protein, Biochemistry 30, 7369–7372 (1991).

  17. 17

    Chen, Y. et al. Three dimensional structure of echistatin and dynamics of the active site. J. biomotec. NMR 4, 307–324 (1994).

  18. 18

    Senn, H. & Klaus, W. The nuclear magnetic resonance structure of flavoridin, an antagonist of the platelet Gp IIb-IIIa receptor, J. molec. Biol. 232, 907–925 (1993).

  19. 19

    Jaseja, M. et al. 1H-NMR studies and secondary structure of the RGD-containing snake toxin, albolabrin, Eur. J. Biochem 218, 853–860 (1993).

  20. 20

    Krezel, A.M., Wagner, G., Seymour-Ulmer, J. & Lazarus, R.A. Structure of the RGD protein decorsin: conserved motif and distinct function in leech proteins that affect blood clotting, Science 264, 1944–1947 (1994).

  21. 21

    Lee, G. et al. Strong inhibition of fibrinogen binding to platelet receptor αIIbβ3 by RGD sequences installed into a presentation scaffold, Prot. Engng. 6, 745–754 (1993).

  22. 22

    Knapp, A., Degenhardt, T. & Dodt, J., Hirudisins, hirudin-derived thrombin inhibitors with disintegrin activity, J. biol. Chem. 267, 24,230–24,234 (1992).

  23. 23

    Dufourcq, J. et al. Structure function relationships for cardiotoxins interacting with phopholipids, Toxicon 20, 165–174 (1982).

  24. 24

    Le Du, M.H., Marchot, P., Bougis, P.E. & Fontecilla-Camps, J.C. 1.9 Å resolution structure of fasciculin 1, an anti-acetylcholinesterase toxin from green mamba snake venom, J. biol. Chem. 267, 22,122–22,130 (1992).

  25. 25

    Havel, T.F. An evaluation of computational strategies for use in the determination of protein structure from distance constraints obtained by nuclear magnetic resonance. Prog. Biophys. molec. Biol. 56, 43–78 (1991).

  26. 26

    Kabsch, W. & Sander, C. Dictionary of protein secondary structure; pattern recognition of hydrogen bonded and geometrical features, Biopolymers 22, 2577–2637 (1983).

  27. 27

    Williams, J.A., Rucinski, B., Holt, J. & Niewiarowski, S. Elegantin and albolabrin purified peptides from viper venoms; homologies with the RGDS domain of fibrinogen and von Willebrand factor, Biochim. biophys. Acta. 1039, 81–89, (1990).

  28. 28

    Gan, Z.R., Gould, R.J., Jacobs, J.W., Friedman, P.A. & Polokoff, M.A. Echistatin - a potent platelet aggregation inhibitor from the venom of the viper echis carinatus. J. biol. Chem. 263, 19,827–19,832 (1988).

  29. 29

    Calvete, J.J. et al. The disulphide bridge pattern of snake venom disintegrins, flavoridin and echistatin. FEBS lett. 309, 316–320, (1992).

  30. 30

    Strydom, D.J. Snake venom toxins: The amino acid sequences of two toxins from Dendroaspis polylepsis polylepsis (black mamba) venom, J. biol. Chem. 247, 4029–4042, (1972).

  31. 31

    Sato, S. & Tamiya, N. The aminoacids of erabutoxins, neurotoxic proteins of sea-snake (Lauticauda semifasciata) Biochem. J. 122, 453–461, (1971).

  32. 32

    Louw, A.I. Snake venom toxins, the complete amino acid sequence of cytotoxin VII4 from the venom of Naja mossambica mossambica. Biochem. biophys. res. Commun. 58, 1022–1029 (1974).

  33. 33

    Viljoen, C.C. & Botes, D.P. Snake venom toxins - the purification and amino acid sequence of toxin Ta2 from Dendroaspis angusticeps venom. J. biol. Chem. 249, 366–372 (1973).

  34. 34

    Hyberts, S.G. Golberg, M.S., Havel, T.F. & wagner, G. The solution structure of eglin c based on measurements of many NOEs and coupling constants and its comparison with X-ray structures. Prot. Sci. 1, 736–751 (1992).

  35. 35

    Morris, A.L. MacAurthur, M.W., Hutchinsom, G.E. & Thornton, J.M. Stereochemical quality of protein structure coordinates. Proteins, 12, 345–364 (1992).

Download references

Author information

Author notes

    • Michael J. Sutcliffe

    Present address: Department of Chemistry, University of Leicester, Leicester, LE1 7RH, U.K.

Affiliations

  1. Biological NMR Centre, University of Leicester, P.O.Box 138, Leicester, LE1 9HN, U.K.

    • Michael J. Sutcliffe
  2. School of Biochemistry, University of Birmingham, Edgbaston, Birmingham, B15 2TT, U.K.

    • Mahesh Jaseja
    •  & Eva I. Hyde
  3. Thrombosis Research Institute, Emmanuel Kaye Building, Manresa Rd., London, SW3 6LR, U.K.

    • Xinjie Lu
  4. Department of Applied Pharmacology, National Heart and Lung Institute, Dovehouse Street, London, SW3 6LY, U.K.

    • Janice A. Williams

Authors

  1. Search for Michael J. Sutcliffe in:

  2. Search for Mahesh Jaseja in:

  3. Search for Eva I. Hyde in:

  4. Search for Xinjie Lu in:

  5. Search for Janice A. Williams in:

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/nsb1194-802