Purification, crystallization and preliminary X-ray diffraction analysis of the glyoxalase II from Leishmania infantum.
In trypanosomatids, trypanothione replaces glutathione in all glutathione-dependent processes. Of the two enzymes involved in the glyoxalase pathway, glyoxalase I and glyoxalase II, the latter shows absolute specificity towards trypanothione thioester, making this enzyme an excellent model to understand the molecular basis of trypanothione binding. Cloned glyoxalase II from Leishmania infantum was overexpressed in Escherichia coli, purified and crystallized. Crystals belong to space group C222(1) (unit-cell parameters a = 65.6, b = 88.3, c = 85.2 angstroms) and diffract beyond 2.15 angstroms using synchrotron radiation. The structure was solved by molecular replacement using the human glyoxalase II structure as a search model. These results, together with future detailed kinetic characterization using lactoyltrypanothione, should shed light on the evolutionary selection of trypanothione instead of glutathione by trypanosomatids.
Trincão
José
J
REQUIMTE-CQFB, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
Sousa Silva
Marta
M
Barata
Lídia
L
Bonifácio
Cecília
C
Carvalho
Sandra
S
Tomás
Ana Maria
AM
Ferreira
António E N
AE
Cordeiro
Carlos
C
Ponces Freire
Ana
A
Romão
Maria João
MJ
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D018314:Leishmania infantum / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013869:Thiolester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction; D000099301:Hydroxyacylglutathione Hydrolase
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D013869:Thiolester Hydrolases; D000099301:Hydroxyacylglutathione Hydrolase