Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Autor(es)

Feliciano, Patricia R. ^[1] ; Gupta, Shreedhara ^[2] ; Dyszy, Fabio ^[3] ; Dias-Baruffi, Marcelo ^[4] ; Costa-Filho, Antonio J. ^[5] ; Michels, Paul A. M. ^[2] ; Nonato, M. Cristina ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, FCFRP, Lab Cristalog Prot, BR-14040903 Ribeirao Preto - Brazil

^[2] Catholic Univ Louvain, Duve Inst & Lab Biochem, Trop Dis Res Unit, B-1200 Brussels - Belgium

^[3] Univ Sao Paulo, IFSC, Grp Biofis Mol Sergio Mascarenhas, BR-13560970 Sao Carlos, SP - Brazil

^[4] Univ Sao Paulo, FCFRP, Lab Glicoimunobiol, BR-14040903 Ribeirao Preto - Brazil

^[5] Univ Sao Paulo, FFCLRP, Dept Fis, Lab Biofis Mol, BR-14040901 Ribeirao Preto - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 51, n. 1-2, p. 25-31, JUL-AUG 2012.
• Citações Web of Science: 14

Resumo

Fumarate hydratases (FHs; EC 4.2.1.2) are enzymes that catalyze the reversible hydration of fumarate to S-malate. Parasitic protists that belong to the genus Leishmania and are responsible for a complex of vector-borne diseases named leishmaniases possess two genes that encode distinct putative FH enzymes. Genome sequence analysis of Leishmania major Friedlin reveals the existence of genes LmjF24.0320 and LmjF29.1960 encoding the putative enzymes LmFH-1 and LmFH-2, respectively. In the present work, the FH activity of both L. major enzymes has been confirmed. Circular dichroism studies suggest important differences in terms of secondary structure content when comparing LmFH isoforms and even larger differences when comparing them to the homologous human enzyme. CD melting experiments revealed that both LmFH isoforms are thermolabile enzymes. The catalytic efficiency under aerobic and anaerobic environments suggests that they are both highly sensitive to oxidation and damaged by oxygen. Intracellular localization studies located LmFH-1 in the mitochondrion, whereas LmFH-2 was found predominantly in the cytosol with possibly also some in glycosomes. The high degree of sequence conservation in different Leishmania species, together with the relevance of FH activity for the energy metabolism in these parasites suggest that FHs might be exploited as targets for broad-spectrum antileishmanial drugs. (c) 2012 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 08/08262-6 - Caracterização cinética, estrutural e funcional dos genes LmjF24.0320. e LmjF29.1960 que codificam a enzima fumarato hidratase em Leishmania major
• Beneficiário: Maria Cristina Nonato
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 06/05538-5 - Clonagem, expressão heteróloga e caracterização do gene LmjF24.0320 que codifica a enzima fumarato hidratase em Leishmania major
• Beneficiário: Patrícia Rosa Feliciano
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 09/10454-3 - Estudos da correlação entre estrutura e função da enzima fumarato hidratase em Leishmania major
• Beneficiário: Patrícia Rosa Feliciano
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 09/15810-2 - Marcação de Spin Sítio Dirigida e Ressonância Magnética Eletrônica: Uma nova abordagem para o estudo de interações membrana-proteína e proteína-proteína
• Beneficiário: Fábio Henrique Dyszy
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado