Rhodocetin- -induced Neuropilin-1-cMet Association Triggers Restructuring of Matrix Contacts in Endothelial Cells

S. Niland, B. Ditkowski, D. Parrandier, L. Roth, H. Augustin, J. A. Eble
2013 Arteriosclerosis, Thrombosis and Vascular Biology  
L.R., H.A.). The online-only Data Supplement is available with this article at http://atvb.ahajournals.org/lookup/suppl/Objective-The snake venom component rhodocetin-αβ (RCαβ) stimulates endothelial cell motility in an α2β1 integrinindependent manner. We aimed to elucidate its cellular and molecular mechanisms. Methods and Results-We identified neuropilin-1 (Nrp1) as a novel target of RCαβ by protein-chemical methods. RCαβ and vascular endothelial growth factor (VEGF)-A avidly bind to Nrp1.
more » ... tead of acting as VEGF receptor 2 coreceptor, Nrp1 associates upon RCαβ treatment with cMet. Furthermore, cell-based ELISAs and kinase inhibitor studies showed that RCαβ induces phosphorylation of tyrosines 1234/1235 and thus activation of cMet. Consequently, paxillin is phosphorylated at Y31, which is redistributed from streak-like focal adhesions to spot-like focal contacts at the cell perimeter, along with α2β1 integrin, thereby regulating cell-matrix interactions. Cortactin is abundant in the cell perimeter, where it is involved in the branching of the cortical actin network of lamellipodia, whereas tensile force-bearing actin stress fibers radiating from focal adhesions disappear together with zyxin, a focal adhesion marker, on RCαβ treatment. Conclusion-Our data demonstrate that (1) Nrp1 is a novel target for venom components, such as RCαβ; (2) Nrp1 coupled to cMet regulates the type of cell-matrix interactions in a manner involving paxillin phosphorylation; and (3) altered cell-matrix interactions determine endothelial cell migration and cellular force management. (Arterioscler Thromb Vasc Biol. 2013;33:544-554.)
doi:10.1161/atvbaha.112.00006 pmid:23288161 fatcat:wrfkcxdyo5aelbxpv54jkkq4va