An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution

0543494 - MBÚ 2022 RIV GB eng J - Journal Article
Del Amo-Maestro, L. - Sagar, A. - Pompach, Petr - Goulas, T. - Scavenius, C. - Ferrero, D. S. - Castrillo-Briceno, M. - Taules, M. - Enghild, J. J. - Bernado, P. - Gomis-Ruth, F.X.
An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution.
Journal of Molecular Biology. Roč. 433, č. 13 (2021), č. článku 166954. ISSN 0022-2836. E-ISSN 1089-8638
R&D Projects: GA MŠMT(CZ) ED1.1.00/02.0109
EU Projects: European Commission(XE) 823839 - EPIC-XS
Research Infrastructure: CIISB II - 90127
Institutional support: RVO:61388971
Keywords : protein-protein interactions * metallopeptidase * biophysical techniques * platelet aggregation * blood coagulation
OECD category: Biochemistry and molecular biology
Impact factor: 6.151, year: 2021
Method of publishing: Open access
https://www.sciencedirect.com/science/article/pii/S0022283621001558

Von Willebrand Factor (vWF), a 300-kDa plasma protein key to homeostasis, is cleaved at a single site by multi-domain metallopeptidase ADAMTS-13. vWF is the only known substrate of this peptidase, which circulates in a latent form and becomes allosterically activated by substrate binding. Herein, we characterised the complex formed by a competent peptidase construct (AD13-MDTCS) comprising metallopeptidase (M), disintegrin-like (D), thrombospondin (T), cysteine-rich (C), and spacer (S) domains, with a 73-residue functionally relevant vWF-peptide, using nine complementary techniques. Pull-down assays, gel electrophoresis, and surface plasmon resonance revealed tight binding with submicromolar affinity. Cross-linking mass spectrometry with four reagents showed that, within the peptidase, domain D approaches M, C, and S. S is positioned close to M and C, and the peptide contacts all domains. Hydrogen/deuterium exchange mass spectrometry revealed strong and weak protection for C/D and M/S, respectively. Structural analysis by multi-angle laser light scattering and small-angle X-ray scattering in solution revealed that the enzyme adopted highly flexible unbound, latent structures and peptide-bound, active structures that differed from the AD13-MDTCS crystal structure. Moreover, the peptide behaved like a self-avoiding random chain. We integrated the results with computational approaches, derived an ensemble of structures that collectively satisfied all experimental restraints, and discussed the functional implications. The interaction conforms to a 'fuzzy complex' that follows a 'dynamic zipper' mechanism involving numerous reversible, weak but additive interactions that result in strong binding and cleavage. Our findings contribute to illuminating the biochemistry of the vWF:ADAMTS-13 axis. (C) 2021 Elsevier Ltd. All rights reserved.
Permanent Link: http://hdl.handle.net/11104/0320685