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Allosteric activation of ADAMTS13 by von Willebrand factor

Joshua Muia1, Jian Zhu1, Garima Gupta1

  • 1Departments of Medicine.

Proceedings of the National Academy of Sciences of the United States of America
|December 17, 2014
PubMed

Insights

The metalloprotease ADAMTS13 regulates blood clotting by cleaving von Willebrand factor (VWF). Its activity is controlled by substrate binding, which relieves inhibition by distal domains, optimizing VWF cleavage under shear stress.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Hematology

Background:

  • ADAMTS13 is a metalloprotease crucial for preventing thrombotic microangiopathies by cleaving von Willebrand factor (VWF).
  • ADAMTS13 deficiency leads to severe microvascular thrombosis due to uncleaved VWF.
  • The enzyme comprises metalloprotease (M), disintegrin-like (D), thrombospondin-1 (T), Cys-rich (C), and spacer (S) domains (MDTCS), along with C-terminal T and CUB domains.

Purpose of the Study:

  • To investigate the functional role of the C-terminal domains of ADAMTS13.
  • To elucidate the regulatory mechanism of ADAMTS13 activity.
  • To understand how substrate binding influences ADAMTS13's interaction with von Willebrand factor.

Main Methods:

  • Small angle X-ray scattering (SAXS) to determine the structure of ADAMTS13.
  • Biochemical assays to measure substrate cleavage activity.
  • Binding studies using VWF and monoclonal antibodies.

Main Results:

  • The distal T8-CUB2 domains of ADAMTS13 were found to significantly inhibit its protease activity.
  • Binding of VWF or specific antibodies to the distal domains relieved this autoinhibition.
  • SAXS data revealed interactions between the distal T-CUB domains and the proximal MDTCS domains.

Conclusions:

  • ADAMTS13 activity is regulated by substrate-induced allosteric activation.
  • This mechanism optimizes VWF cleavage under physiological conditions, such as fluid shear stress.
  • Similar allosteric regulatory mechanisms may exist in other ADAMTS family proteases.
Keywords:
allosteric regulationhemostasismetalloproteases

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