1. Identification and validation of Trypanosoma cruzi's glycosomal adenylate kinase containing a peroxisomal targeting signal.
- Author
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Milagros Camara Mde L, Bouvier LA, Miranda MR, and Pereira CA
- Subjects
- 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, Adenylate Kinase metabolism, Microbodies enzymology, Peroxisomes enzymology, Signal Transduction, Trypanosoma cruzi enzymology
- 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., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
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