title: Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of glyoxalase I from Leishmania infantum.
abstract: Glyoxalase I (GLO1) is the first of the two glyoxalase-pathway enzymes. It catalyzes the formation of S-D-lactoyltrypanothione from the non-enzymatically formed hemithioacetal of methylglyoxal and reduced trypanothione. In order to understand its substrate binding and catalytic mechanism, GLO1 from Leishmania infantum was cloned, overexpressed in Escherichia coli, purified and crystallized. Two crystal forms were obtained: a cube-shaped form and a rod-shaped form. While the cube-shaped form did not diffract X-rays at all, the rod-shaped form exhibited diffraction to about 2.0 A resolution. The crystals belonged to space group P2(1)2(1)2, with unit-cell parameters a = 130.03, b = 148.51, c = 50.63 A and three dimers of the enzyme per asymmetric unit.
authors.affiliation: Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Lisboa, Campo Grande, Edificio C8, 1149-016 Lisboa, Portugal.
authors.forename: Lídia
authors.initials: L
authors.lastname: Barata
authors.forename: Marta
authors.initials: M
authors.lastname: Sousa Silva
authors.forename: Linda
authors.initials: L
authors.lastname: Schuldt
authors.forename: Gonçalo
authors.initials: G
authors.lastname: da Costa
authors.forename: Ana M
authors.initials: AM
authors.lastname: Tomás
authors.forename: António E N
authors.initials: AE
authors.lastname: Ferreira
authors.forename: Manfred S
authors.initials: MS
authors.lastname: Weiss
authors.forename: Ana
authors.initials: A
authors.lastname: Ponces Freire
authors.forename: Carlos
authors.initials: C
authors.lastname: Cordeiro
mesh_terms: D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D015870:Gene Expression; D007791:Lactoylglutathione Lyase / Q000737:chemistry* / Q000302:isolation & purification; D018314:Leishmania infantum / Q000201:enzymology*
chemical_list: D007791:Lactoylglutathione Lyase
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