RRC ID 65884
Author Seo T, Sakon T, Nakazawa S, Nishioka A, Watanabe K, Matsumoto K, Akasaka M, Shioi N, Sawada H, Araki S.
Title Haemorrhagic snake venom metalloproteases and human ADAMs cleave LRP5/6, which disrupts cell-cell adhesions in vitro and induces haemorrhage in vivo.
Journal FEBS J
Abstract Snake venom metalloproteases (SVMPs) are members of the a disintegrin and metalloprotease (ADAM) family of proteins, as they possess similar domains. SVMPs are known to elicit snake venom-induced haemorrhage; however, the target proteins and cleavage sites are not known. In this work, we identified a target protein of vascular apoptosis-inducing protein 1 (VAP1), an SVMP, relevant to its ability to induce haemorrhage. VAP1 disrupted cell-cell adhesions by relocating VE-cadherin and γ-catenin from the cell-cell junction to the cytosol, without inducing proteolysis of VE-cadherin. The Wnt receptors low-density lipoprotein receptor-related proteins 5 and 6 (LRP5/6) are known to promote catenin relocation, and are rendered constitutively active in Wnt signalling by truncation. Thus, we examined whether VAP1 cleaves LRP5/6 to induce catenin relocation. Indeed, we found that VAP1 cleaved the extracellular region of LRP6 and LRP5. This cleavage removes four inhibitory β-propeller structures, resulting in activation of LRP5/6. Recombinant human ADAM8 and ADAM12 also cleaved LRP6 at the same site. An antibody against a peptide including the LRP6-cleavage site inhibited VAP1-induced VE-cadherin relocation and disruption of cell-cell adhesions in cultured cells, and blocked haemorrhage in mice in vivo. Intriguingly, animals resistant to the effects of haemorrhagic snake venom express variants of LRP5/6 that lack the VAP1-cleavage site, or low-density lipoprotein receptor domain class A domains involved in formation of the constitutively active form. The results validate LRP5/6 as physiological targets of ADAMs. Furthermore, they indicate that SVMP-induced cleavage of LRP5/6 causes disruption of cell-cell adhesion and haemorrhage, potentially opening new avenues for the treatment of snake bites.
Volume 284(11)
Pages 1657-1671
Published 2017-6-1
DOI 10.1111/febs.14066
PMID 28425175
MeSH ADAM Proteins / metabolism* ADAM Proteins / pharmacology ADAM12 Protein / metabolism ADAM12 Protein / pharmacology Amino Acid Sequence Animals Antibodies, Neutralizing / pharmacology Apoptosis Regulatory Proteins / metabolism* Cell Adhesion / drug effects Cell Adhesion / physiology Crotalid Venoms / metabolism* Drug Resistance Fibrinogen / metabolism Fibronectins / metabolism HeLa Cells Hemorrhage / chemically induced* Human Umbilical Vein Endothelial Cells Humans Hydrophobic and Hydrophilic Interactions Low Density Lipoprotein Receptor-Related Protein-5 / chemistry Low Density Lipoprotein Receptor-Related Protein-5 / physiology* Low Density Lipoprotein Receptor-Related Protein-6 / chemistry Low Density Lipoprotein Receptor-Related Protein-6 / physiology* Male Membrane Proteins / metabolism Membrane Proteins / pharmacology Metalloendopeptidases / metabolism* Mice Models, Molecular Molecular Docking Simulation Protein Domains Protein Structure, Secondary / drug effects Recombinant Proteins / metabolism Sequence Alignment Sequence Homology, Amino Acid Species Specificity Vertebrates / metabolism Wnt Signaling Pathway / drug effects Wnt Signaling Pathway / physiology
IF 4.392
Human and Animal Cells HeLa