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Expression, purification, activity assay and crystallization of the enzyme UCMP kinase from biosynthesis of pyrimidine nucleotides of the parasite Schistosoma mansoni

Grant number: 14/15594-6
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): September 01, 2014
Effective date (End): June 30, 2015
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Humberto D'Muniz Pereira
Grantee:Marilia Mendes Raddi
Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Neglected tropical diseases are considered a major public health problem, being mainly 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 requires the assistance of DNA bases for your metabolism and requires an active metabolism of nucleotides. Unlike the metabolism of purines, which contains only the salvation pathway to meet the demand of purine bases, the pyrimidine metabolism pathway has both "de novo" as well as pyrimidine salvation pathway. 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 in S. mansoni, uridylate/cytidylate kinase (UCMP kinase). 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 its 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.

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