title: Molecular and biochemical characterisation of Trypanosoma cruzi phosphofructokinase.
abstract: The characterisation of the gene encoding Trypanosoma cruzi CL Brener phosphofructokinase (PFK) and the biochemical properties of the expressed enzyme are reported here. In contradiction with previous reports, the PFK genes of CL Brener and YBM strain T. cruzi were found to be similar to their Leishmania mexicana and Trypanosoma brucei homologs in terms of both kinetic properties and size, with open reading frames encoding polypeptides with a deduced molecular mass of 53,483. The predicted amino acid sequence contains the C-terminal glycosome-targeting tripeptide SKL; this localisation was confirmed by immunofluorescence assays. In sequence comparisons with the genes of other eukaryotes, it was found that, despite being an adenosine triphosphate-dependent enzyme, T. cruzi PFK shows significant sequence similarity with inorganic pyrophosphate-dependent PFKs.
authors.affiliation: Centro de Biotecnología, Instituto de Estudios Avanzados, Caracas, Venezuela.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Rodríguez
authors.forename: Noelia
authors.initials: N
authors.lastname: Lander
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Ramirez
mesh_terms: D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000032:analysis*; D008856:Microscopy, Fluorescence; D025221:Phosphofructokinases / Q000235:genetics*; D013329:Structure-Activity Relationship; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics
chemical_list: D016054:DNA, Protozoan; D025221:Phosphofructokinases