Immobilization nucleoside diphosphate kinase (NDK) enzyme Leishmania major: the study of conditions for the application in the screening of ligands

• Grant number: 14/06907-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: July 01, 2014
• End date: January 31, 2018
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Carmen Lúcia Cardoso
• Grantee: Juliana Maria de Lima
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 13/01710-1 - Enzyme ligand: new models of screening, AP.TEM

Abstract

According to the World Health Organization (WHO), leishmaniasis is a neglected disease that afflicts people in underdeveloped countries, and disfigures, stigmatizes and kills. Therefore, through a Special Research and Training Programme (TDR) the WHO aimed to eradicate this disease, which is currently treated with expensive and toxic drugs. Researchers have characterized and purified proteins of microorganisms that are pathogenic to humans, a strategy in the search for molecules that can act as biological targets. The enzyme Nucleoside Diphosphate Kinase B (NDKb) has a key role in maintaining the amount of intracellular NTPs and dNTPs it is secreted by Leishmania major and is determines the invasive / evasive process during successful infection of macrophages. Considering that the enzyme is a potential biological target, it is necessary to develop and/or upgrade techniques, to conduct skillful studies on this biomolecule. Biochromatography has successfully been used in this sense and the association of IMERs with chromatographic systems is a highly efficient multidimensional method that is potentially applicable in the screening of selective inhibitors. Hence, in this project we propose the immobilization of enzymes NDKb of Leishmania major and its mutant K30A onto silica capillary, which will later be engaged in a multidimensional method that will be developed and validated. (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LOPES VILELA, ADRIANA FERREIRA; SEIDL, CLAUDIA; LIMA, JULIANA MARIA; CARDOSO, CARMEN LUCIA. An improved immobilized enzyme reactor-mass spectrometry-based label free assay for butyrylcholinesterase ligand screening. Analytical Biochemistry, v. 549, p. 53-57, MAY 15 2018. (16/02873-0, 14/11640-3, 14/06907-0)

SEIDL, CLAUDIA; DE LIMA, JULIANA MARIA; LEME, GABRIEL MAZZI; PIRES, ANANDA FERREIRA; STOLL, DWIGHT R.; CARDOSO, CARMEN LUCIA. A Comprehensive 2D-LC/MS Online Platform for Screening of Acetylcholinesterase Inhibitors. FRONTIERS IN MOLECULAR BIOSCIENCES, v. 9, p. 14-pg., 2022-03-16. (14/50249-8, 14/11640-3, 14/06907-0, 19/05363-0, 16/02941-5)

LIMA, JULIANA M.; SEIDL, CLAUDIA; CUNHA, ELISE M. F.; OLIVEIRA, ARTHUR H. C.; CARDOSO, CARMEN L.. On-Flow Ligand Screening Assay Based on Immobilized Nucleoside Diphosphate Kinase B from Homo sapiens. Journal of the Brazilian Chemical Society, v. 30, n. 11, SI, p. 2308-2317, NOV 2019. (14/50249-8, 13/01710-1, 14/06907-0, 14/11640-3)

LIMA, JULIANA MARIA; VIEIRA, PLINIO SALMAZO; CAVALCANTE DE OLIVEIRA, ARTHUR HENRIQUE; CARDOSO, CARMEN LUCIA. Label-free offline versus online activity methods for nucleoside diphosphate kinase b using high performance liquid chromatography. ANALYST, v. 141, n. 15, p. 4733-4741, 2016. (14/06907-0, 13/01710-1, 11/20569-2)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

LIMA, Juliana Maria de. Enzymatic on-line assay based on immobilized enzyme coupled to zonal chromatography to identify and characterize inhibitors for Nucleoside Diphosphate Kinase B. 2018. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC) Ribeirão Preto.