Planejamento racional de drogas contra tripanosomatídeos: gGAPDH de Trypanosoma cruzi e XPRT de Leishmania major / Rational design of anti-trypanosomatids drugs: T. cruzi gGAPDH and Leishmania major XPRT

AUTOR(ES)
DATA DE PUBLICAÇÃO

2004

RESUMO

Aiming at discover molecules with good inhibitory activity against tripanosomatides enzymatic targets, the crystallographic structures of glyceraldehydes-3-phosphate dehydrogenase in complex with 1,3 bisfosfoglyceric acid analogues (30 and 33)were solved, molecular modeling studies were undertaken and xprt (xanthine phosphorybosil transferase) gene from Leishmania major was cloned and over-expressed in Escherichia coli. The enzyme thus obtained was purified and kinetically characterized. .gGAPDH-33 complex, up to 2,5A resolution revealed the tioketal intermediate binding mode. The final model was refined to R 0.20 from a 97,5% completeness dataset. The crystallographic structure gives, for the first time, experimental evidence for the flip-flop mechanism, which describes how the substrate goes from inorganic-phosphate binding site to substratephosphate binding site. gGAPDH-30 complex, solved to 2,75A resolution, revealed the inhibitor binding mode. The final model has R= 0,19 and was refined from a 92,4% completeness dataset. This structure was used as the framework upon which modeling studies were performed. Modeling results suggest why these inhibitors show a different inhibitory profile against Trypanosoma brucei and Trypanosoma cruzi. L. major XPRT shows a high affinity for hypoxanthine, an alternative substrate, when compared to L. donovani XPRT, aiming at understand this behavior homology modeling studies are currently under progress.

ASSUNTO(S)

cristalografia de raios x ggapdh x-ray crystallography trypanosomatids xprt ggapdh docking docking tripanosomatídeos xprt

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