Notícies
20.04.2021
Publicació de l'article "An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution "
La Dra. Marta Taulés (Anàlisi Interaccions Moleculars dels CCITUB), ha participat en la publicació de l'article "An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution" a la revista Journal of Molecular Biology en col·laboració amb investigadors/es del CSIC, de la Université de Montpellier, de l’Institute of Microbiology of the Czech Academy of Sciences i de la Aarhus University
A continuació es mostren els aspectes més destacats de l’article:
• El factor Von Willebrand (vWF) es divideix en un sol lloc per la peptidasa ADAMTS-13.
• Es va caracteritzar el complex entre un pèptid vWF i ADAMTS-13 mitjançant nou tècniques.
• La interacció s’ajusta a un complex difús que segueix un mecanisme dinàmic de cremallera.
• Moltes interaccions reversibles, febles però additives donen lloc a una forta unió i escissió.
El resum de l'article és el següent:
"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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