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Exp Parasitol ; 130(4): 408-11, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22343032

ABSTRACT

Adenylate kinases are key enzymes involved in cell energy management. Trypanosomatid organisms have the largest number of isoforms found in a single cell, constituting a major difference with the mammalian hosts. In this work we study an adenylate kinase, TcADK3, the only Trypanosoma cruzi protein harboring the putative peroxisomal (glycosomal) targeting signal, "-CKL". Parasites expressing GFP fused to TcADK3 showed a strong fluorescence in the glycosomes. The same result was obtained when the tripeptide "-CKL" was added at the C-terminus of the GFP, demonstrating that this signal is necessary and sufficient for targeting proteins to glycosomes. When this tripeptide was removed from the GFP-TcADK3 fusion protein, the fluorescence was re-localized in the cytoplasm. The CKL signal could be used for targeting foreign proteins to the glycosomes. This model also provides a useful tool to study glycosomes dynamics, morphology or number in living parasites in any stage of the life cycle.


Subject(s)
Adenylate Kinase/metabolism , Microbodies/enzymology , Peroxisomes/enzymology , Signal Transduction , Trypanosoma cruzi/enzymology , Adenylate Kinase/chemistry , Adenylate Kinase/genetics , Amino Acid Sequence , Cloning, Molecular , Gene Expression Regulation, Enzymologic , Isoenzymes/chemistry , Isoenzymes/genetics , Isoenzymes/metabolism , Microscopy, Fluorescence , Plasmids , Transfection , Trypanosoma cruzi/genetics
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