Functional and molecular characterization of a glycosomal PPi-dependent enzyme in trypanosomatids: Pyruvate, phosphate dikinase
| Autor: | Frédéric Bringaud, D. Baltz, Théo Baltz |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Multidisciplinary
biology Molecular Sequence Data Trypanosoma brucei brucei Protozoan Proteins Trypanosoma brucei Biological Sciences biology.organism_classification Glycosome Trypanosoma vivax Pyruvate Orthophosphate Dikinase Enzyme Activation Pyruvate phosphate dikinase Enzyme activator Biochemistry Crithidia parasitic diseases Escherichia coli Animals Glycolysis Amino Acid Sequence Cloning Molecular Trypanosoma cruzi |
| Popis: | Trypanosomatids are parasitic protists that have an ATP-dependent glycolysis with no indication of PP i -dependent metabolism. Most of the glycolysis takes place in peroxisome-like organelles, the glycosomes. We characterized in Trypanosoma brucei a single-copy gene encoding a PP i -dependent enzyme, pyruvate, phosphate dikinase (PPDK), which was expressed functionally in Escherichia coli . Specific antibodies detected a 100-kDa protein in procyclic forms but not in mammalian forms of T. brucei , indicating a differential expression. Glycosomal localization of PPDK was determined by immunofluorescence analysis and was confirmed by Western blot analysis on glycosomal fractions by using anti-PPDK antibodies. Expression and localization of recombinant PPDKs in procyclic forms of T. brucei showed that the AKL motif at the C-terminal extremity of PPDK is necessary for glycosomal targeting. PPDK was detected in every trypanosomatid tested— Trypanosoma congolense , Trypanosoma vivax , Trypanosoma cruzi , Phytomonas , Crithidia and Leishmania —with a good correlation between amount of protein and enzymatic activity. The precise role of PPDK in trypanosomatid carbohydrate metabolism remains to be clarified. |
| Databáze: | OpenAIRE |
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