Entamoeba histolytica, heterologous expression and in situ immunolocalization of ornithine decarboxylase (EhODC). Experimental parasitology [Exp Parasitol] Journal article | | Title | Entamoeba histolytica, heterologous expression and in situ immunolocalization of ornithine decarboxylase (EhODC). | | Author(s) | Terán-Figueroa Y, Arteaga-Nieto P, Bethancourt-Rodrı Guez A, Labra-Barrios ML, Luna-Arias JP, Flores-Carreón A, Cano-Canchola C, López-Romero E, Calvo-Méndez C | | Institution | Instituto de Investigación en Biologi a Experimental, Facultad de Qui mica, Universidad de Guanajuato, Apartado Postal No. 187, Guanajuato, Gto. 36000, México. | | Source | Exp Parasitol 2009 Jun 8. | | Abstract | Previous studies from this laboratory have dealt with the purification and biochemical characterization of ornithine decarboxylase (ODC) from Entamoeba histolytica. Enzyme compartmentalization has been described as a major mechanism in the regulation of polyamine metabolism. However, the subcellular location of ODC in the human parasite has remained unresolved. To examine this issue, we cloned the full-length gene (Ehodc) encoding for the parasite enzyme, whose open reading frame encodes for a peptide of 412 amino acids with an estimated molecular mass of 46 kDa that exhibits similarity to other ODCs. Heterologous overexpression of the gene allowed us to purify the recombinant protein (rEhODC) by metal affinity chromatography. The purified polypeptide was used to raise heteroclonal antibodies that were utilized to localize the enzyme in situ by immunofluorescence and confocal microscopy. EhODC was observed to be associated with the plasma membrane, in vesicles close to the plasma membrane and in the EhkOs organelle. | | Language | ENG | | Pub Type(s) | JOURNAL ARTICLE
| | PubMed ID | 19520076 |
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