Immobilization studies for arginase

• Grant number: 17/14046-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2017
• End date: August 31, 2018
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Quezia Bezerra Cass
• Grantee: Adriana Arnosti Bonatti
• Host Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Enzymes have become important targets in the search for inhibitors as a strategy in the search for new drugs. The process of discovery of new ligands begins by the search for compounds that modulate the activity of vital enzymes to the pathogen. Thus, together, it becomes necessary to develop new models of screening for hits as drug candidates. Arginase is a metalloenzyme of L. amazonenses, a causative agent of leishmaniasis, which catalyzes the hydrolysis of L-arginine into L-ornithine and urea, the first reaction in the metabolic pathway of polyamines, essential molecules in most living organisms. Because of its importance in the metabolism of the parasite, the enzyme has become a promising target for the development of new drugs against leishmaniasis. Conventional methods for the determination of the catalytic activity of arginase use unnatural substrates, isotopically labeled reagents or coupled to the enzymatic reaction of urease/glutamate dehydrogenase, with the monitoring of NADH consumed. In coupled assays, any interference in the activity of the second enzyme may produce false-positive results. The use of immobilized enzyme reactors (IMERs) provides several advantages over conventional assays, which include increasing the life and stability of the enzyme, the possibility of reusing the protein and the greater reproducibility of the data. Thus, conditions for arginase immobilization and modulation of assays for the IMERs produced is the purpose of the project proposed here. (AU)