Recombinant production of TGFb cytokine for the analysis of its proteolytic processing

• Grant number: 19/24805-4
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Start date: February 01, 2020
• End date: March 22, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: André Zelanis Palitot Pereira
• Grantee: Uilla Barcick
• Supervisor: Nicholas J. Gay
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Federal de São Paulo (UNIFESP). Campus São José dos Campos. São José dos Campos , SP, Brazil
• Institution abroad: University of Cambridge, England
• Associated to the scholarship: 19/10817-0 - Evaluation of the proteolytic processing of TGFB citokyne in melanoma secretomes, BP.IC

Abstract

The search for cancer biomarkers is a field in which a large amount of human and financial resources is concentrated, both from private and public research around the world. Experimental approaches in proteomics related to the prospection of biomarkers aim to find specific proteins, characteristic expression patterns, or markers of tumor progression. In a recent study on the proteolytic processing of a paired set of murine melanoma composed of the Melan-a (a normal melanocyte) and Tm1 (the corresponding tumor phenotype) cell lines, it was possible to identify, exclusively in the tumor phenotype, proteolytic processing events in proteins of important role in tumor development. Among the cleavage sites observed, we identified a peptide corresponding to the processing of the TGFb-3 precursor protein in a portion immediately prior to the canonical cleavage site for this cytokine. From the functional point of view, this finding suggests a non-canonical and autocrine activation of TGFb-3 by transformed melanocytes. Thus, this project has as main objective to produce the TGFb cytokine in a recombinant form in human embryonic kidney cells (HEK293) aiming to use the recombinant protein for study of its proteolytic processing in a cellular model of murine melanoma. (AU)