Characterization of the inhibition potential of collagen receptor Discoiding domain receptor 2 by snake venom metalloproteases components

• Grant number: 14/01534-1
• Support Opportunities: Scholarships abroad - Research
• Start date: July 01, 2014
• End date: August 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Leo Kei Iwai
• Grantee: Leo Kei Iwai
• Host Investigator: Paul Huang
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Institution abroad: Institute of Cancer Research (ICR), England
• Associated research grant: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling, AP.CEPID

Abstract

Snake venom metalloproteases and disintegrins (SVMPs) are a class of proteins from snake venom presenting several biological functions such as inhibition of platelet aggregation and integrin-dependent cell adhesion by binding to collagen and integrin through different and independent motifs. Similarly to integrins, Discoidin Domain Receptors (DDRs) are known to bind and be activated by collagen. In addition, it has been shown that DDRs increase metalloproteinases secretion and collagen rearrangement. DDRs are a member of tyrosine kinase receptors and much attention has recently been given to this receptor due to the recent discoveries of its potential implication in several diseases including cancer. Although some works has been able to characterize the binding properties of this receptor to collagen and map some of the downstream component of the activation network, still little is known about the mechanisms and function of this receptor. In this work we propose to test the potential DDR2 inhibition by two different SVMP components with differential hemorrhagic properties: HF3 and Bothropasin, as well as the hyper-variable region (HVR) of the cysteine-rich domain of HF3, which is known to present hemorrhagic properties without the disintegrin domain. Effects on several cancer cell lines overexpressing DDR2 molecules will be monitored, where receptor downstream components phosphorylation will be characterized by mass spectrometry-based selective reaction monitoring (SRM) and receptor shedding or internalization behavior will be analyzed by fluorescent microscopy. (AU)