Polyamine biosynthesis in Phytomonas: Biochemical characterisation of a very unstable ornithine decarboxylase

Marcora, María Silvina; Cejas, Silvina; González, Nélida S.; Carrillo, Carolina; Algranati, Israel D.

doi:10.1016/j.ijpara.2010.04.003

Cited by 3 publications

(1 citation statement)

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“…Cell extracts were obtained and the enzymatic activity was measured by the release of 14 CO 2 from L-[1- 14 C]ornithine as previously described [52]. Briefly, parasites were collected at the exponential phase of growth.…”

Section: Methodsmentioning

confidence: 99%

Trypanosoma cruzi Coexpressing Ornithine Decarboxylase and Green Fluorescence Proteins as a Tool to Study the Role of Polyamines in Chagas Disease Pathology

et al. 2011

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Polyamines are essential for Trypanosoma cruzi, the causative agent of Chagas disease. As T. cruzi behaves as a natural auxotrophic organism, it relies on host polyamines biosynthesis. In this paper we obtained a double-transfected T. cruzi parasite that expresses the green fluorescent protein (GFP) and a heterologous ornithine decarboxylase (ODC), used itself as a novel selectable marker. These autotrophic and fluorescent parasites were characterized; the ODC presented an apparent Km for ornithine of 0.51 ± 0.16 mM and an estimated Vmax value of 476.2 nmoles/h/mg of protein. These expressing ODC parasites showed higher metacyclogenesis capacity than the auxotrophic counterpart, supporting the idea that polyamines are engaged in this process. This double-transfected T. cruzi parasite results in a powerful tool—easy to follow by its fluorescence—to study the role of polyamines in Chagas disease pathology and in related processes such as parasite survival, invasion, proliferation, metacyclogenesis, and tissue spreading.

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Section: Methodsmentioning

confidence: 99%