title: Purification, crystallization and preliminary X-ray diffraction analysis of the glyoxalase II from Leishmania infantum.
abstract: 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.
authors.affiliation: REQUIMTE-CQFB, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
authors.forename: José
authors.initials: J
authors.lastname: Trincão
authors.forename: Marta
authors.initials: M
authors.lastname: Sousa Silva
authors.forename: Lídia
authors.initials: L
authors.lastname: Barata
authors.forename: Cecília
authors.initials: C
authors.lastname: Bonifácio
authors.forename: Sandra
authors.initials: S
authors.lastname: Carvalho
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Tomás
authors.forename: António E N
authors.initials: AE
authors.lastname: Ferreira
authors.forename: Carlos
authors.initials: C
authors.lastname: Cordeiro
authors.forename: Ana
authors.initials: A
authors.lastname: Ponces Freire
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Romão
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D013869:Thiolester Hydrolases; D000099301:Hydroxyacylglutathione Hydrolase