Expression, purification, activity assay and crystallization of UCK1 and UCK2 enzymes of the pathway salvation pyrimidines from the parasite Schistosoma mansoni

• Grant number: 14/23659-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2015
• End date: June 30, 2017
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Humberto D'Muniz Pereira
• Grantee: Gabriela Viotto Sarro
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Neglected tropical diseases are considered a public health problem and were primarily related to poverty and low economic development. These neglected diseases account for a major cause of death in developing countries, receiving little or no attention from pharmaceutical companies for the development of therapies. Schistosoma mansoni is the parasite responsible for schistosomiasis. Schistosoma, like every living being, needs the contribution of DNA bases to their metabolism and requires an active metabolism of nucleotides. Unlike the metabolism of purines, which only has the path of salvation to meet the demand of purine bases, this being well reported in the literature, the metabolism of pyrimidines has both via " new " as the way of salvation and pyrimidines scarce literature for these pathways . The objectives of this project are : to express, purify, perform tests of activity and crystallization of one of the enzymes involved in the metabolism of nucleotides of S. mansoni, cytidine uridine kinase 1 and 2 ( and UCK1 UCK2 ) . The gene encoding the enzyme was cloned in high- throughput system together with other enzymes and expression and purification of full-scale tests were performed. The understanding of the structure of this enzyme together with their biochemical activity contributes to the understanding of the parasite and the subsequent identification of key enzymes in this pathway for the development of specific inhibitors.