Noticias
20.04.2021
Publicación del artículo "An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution "
La Dra. Marta Taulés (Análisis de Interacciones Moleculares de los CCITUB), ha participado en la publicación del artículo "An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution" en la revista Journal of Molecular Biology en colaboración con investigadores/as del CSIC, de la Université de Montpellier, del Institute of Microbiology of the Czech Academy of Sciences y de la Aarhus University
A continuación se muestran los aspectos más destacados del artículo:
• El factor Von Willebrand (vWF) se divide en un sólo lugar por la peptidasa ADAMTS-13.
• Se ha caracteritzado el complejo entre un péptido vWF y ADAMTS-13 mediante nueve técnicas.
• La interacción se ajusta a un complejo difuso que sigue un mecanismo dinámico de cremallera.
• Muchas interacciones reversibles, débiles pero additivas dan lugar a una fuerte unión y escisión.
El resumen del artículo es el siguiente:
"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 sub-micromolar 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.""
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