Purification, crystallization and preliminary X-ray diffraction analysis of the glyoxalase II from Leishmania infantum
- REQUIMTE-CQFB, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica (Portugal)
- Centro de Química e Bioquímica, Departamento Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Edifício C8, Lisboa (Portugal)
- IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto (Portugal)
A glyoxalase II from L. infantum was cloned, purified and crystallized and its structure was solved by X-ray crystallography. 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{sub 1} (unit-cell parameters a = 65.6, b = 88.3, c = 85.2 Å) and diffract beyond 2.15 Å 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 trypano-somatids.
- OSTI ID:
- 22360172
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 8; Other Information: PMCID: PMC2242913; PMID: 16880563; PUBLISHER-ID: bw5158; OAI: oai:pubmedcentral.nih.gov:2242913; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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