Open AccessCloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of glyoxalase I from Leishmania infantum
Author(s) -
Barata Lídia,
Sousa Silva Marta,
Schuldt Linda,
Da Costa Gonçalo,
Tomás Ana M.,
Ferreira António E. N.,
Weiss Manfred S.,
Ponces Freire Ana,
Cordeiro Carlos
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110010754
Subject(s) - lactoylglutathione lyase , leishmania infantum , crystallography , methylglyoxal , escherichia coli , chemistry , crystallization , x ray crystallography , leishmania , enzyme , substrate (aquarium) , diffraction , stereochemistry , microbiology and biotechnology , biochemistry , biology , physics , organic chemistry , gene , parasite hosting , optics , world wide web , computer science , ecology
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 Å resolution. The crystals belonged to space group P 2 1 2 1 2, with unit‐cell parameters a = 130.03, b = 148.51, c = 50.63 Å and three dimers of the enzyme per asymmetric unit.