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The snake venom protein botrocetin acts as a biological brace to promote dysfunctional platelet aggregation

Nature Structural & Molecular Biology volume 12, pages 152–159 (2005)Cite this article

Abstract

Botrocetin is a snake venom protein that enhances the affinity of the A1 domain of plasma von Willebrand factor (vWF) for the platelet receptor glycoprotein Ibα (GPIbα), an event that contributes to bleeding and host death. Here we describe a kinetic and crystallographic analysis of this interaction that reveals a novel mechanism of affinity enhancement. Using high-temporal-resolution microscopy, we show that botrocetin decreases the GPIbα off-rate two-fold in both human and mouse complexes without affecting the on-rate. The key to this behavior is that, upon binding of GPIbα to vWF-A1, botrocetin prebound to vWF-A1 makes no contacts initially with GPIbα, but subsequently slides around the A1 surface to form a new interface. This two-step mechanism and flexible coupling may prevent adverse alterations in on-rate of GPIbα for vWF-A1, and permit adaptation to structural differences in GPIbα and vWF in several prey species.

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Figure 1: Structure and comparison of the mouse vWF-A1–botrocetin binary complex.
Figure 2: Structure of the vWF-A1–GPIbα–botrocetin ternary complex.
Figure 3: Structural changes on formation of ternary complex.
Figure 4: Requirement for botrocetin binding to the A1 domain.
Figure 5: The effect of botrocetin on the kinetics of the GPIbα–vWF-A1 tether bond.
Figure 6: The botrocetin-GPIbα interface regulates the off-rate of the ternary complex.
Figure 7

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Acknowledgements

We thank to the staff of the Stanford Synchrotron Radiation Laboratory and the US Department of Energy for the use of X-ray data collection facilities. This work was supported by grants from the US National Institutes of Health (to R.C.L and T.G.D), the American Heart Association (T.G.D.) and a postdoctoral fellowship from the PhRMA Foundation (I.J.L.).

Author information

Author notes

  1. Teresa Doggett and Ian J Laurenzi: These authors contributed equally to this work.

Authors and Affiliations

  1. Infectious and Inflammatory Disease Center, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, 92037, California, USA

    Koichi Fukuda & Robert C Liddington

  2. Department of Pediatrics and Pathology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, 63110, Missouri, USA

    Teresa Doggett & Thomas G Diacovo

  3. Department of Molecular Biophysics and Biochemistry, Yale University, Bass 437, 266 Whitney Avenue, New Haven, 06520, Connecticut, USA

    Ian J Laurenzi

Authors
  1. Koichi Fukuda
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Corresponding authors

Correspondence to Robert C Liddington or Thomas G Diacovo.

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Supplementary information

Supplementary Fig. 1

Intrinsic off-rate for the botrocetin–vWF-A1–GPIb tether bond. (PDF 184 kb)

Supplementary Fig. 2

The effect of botrocetin on the kinetics of the GPIbα–vWF-A1 tether bond. (PDF 330 kb)

Supplementary Fig. 3

The botrocetin-GPIbα interface regulates the off-rate of the ternary complex. (PDF 80 kb)

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Fukuda, K., Doggett, T., Laurenzi, I. et al. The snake venom protein botrocetin acts as a biological brace to promote dysfunctional platelet aggregation. Nat Struct Mol Biol 12, 152–159 (2005). https://doi.org/10.1038/nsmb892

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