Oral Presentation 9th General Meeting of the International Proteolysis Society 2015

A selective pro-angiogenic multifunctional protein extracted from Crataeva tapia bark (#9)

Fabricio P Batista 1 , Yara Lobo 1 , Sheila S Andrade 1 , Maria Tereza Correia 2 , Patricia M.G Paiva 2 , Camila R Bonturi 1 , Carol M Vicente 1 , Leny Toma 1 , Thaysa Paschoalin 1 , Claudia A.A de Paula , Maria Luiza V Oliva 1
  1. Universidade Federal de São Paulo, São Paulo, SãO PAULO, Brazil
  2. Department of Biochemistry, Federal University of Pernambuco, Pernambuco, PE, Brazil

CrataBL, extracted from Crataeva tapia is a multifunctional protein. Besides the lectin properties it inhibits serine protease and binds to glycosaminoglycan. In addition, CrataBL affects the viability of prostate cancer by inducing apoptosis. In this work, the effect of this protein was investigated on human endothelial cells (HUVEC). Our data demonstrate that in addition to its anti-cancer properties, CrataBL selectively stimulated  metabolism of HUVEC in a dose-dependent manner, as well as angiogenic phenotype in vitro, increasing 150% the tube formation with 10 µmol/L. CrataBL also demonstrated a chemoattractant effect to endothelial cells, stimulating migration and invasion by approximately 90%. Furthermore, CrataBL stimulated adhesion to collagen I selectively compared with other adhesion molecules, like collagen IV, fibronectin and laminin. The treatment of endothelial cells with CrataBL showed significant effect in alterations of gelatinases in zimography. The cell signaling investigation demonstrated an increase in both phosphorylation of SRC protein tyrosine kinase, as well as focal adhesion-associated kinase (FAK). It stimulates cell adhesion, cytoskeleton rearrangement, migration, invasion and tube formation. Phosphorylation of the extracellular regulated protein kinase (ERK) and p38-MAPK, that mediated gene expression, proliferation and other angiogenic processes, also were increased. On the other hand, there was reduction on phosphorylation of protein kinase B (AKT/PKB). It is known that, when phosphorylated, AKT/PKB could activate both survival and nitric oxide (NO) liberation. And, in the concentrations tested, CrataBL shows no significant effect on liberation of NO. The investigation of glycosaminoglycan indicated that CrataBL increased remarkably expression of heparan sulfate but not so much of chondroitin sulfate. In conclusion, the effect of CrataBL is triggering FAK-SRC and MAPK pathway, in addition to the expression of glycosaminoglycans, leading to cell and matrix changes that culminate in angiogenesis.