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A Novel Monoclonal Antibody Against the C-terminus of β-Tubulin Recognizes Endocytic Organelles in Trypanosoma cruzi

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Microtubule cytoskeleton is a dynamic structure involved in the maintenance of eukaryote cell shape, motion of cilia and flagellum, and intracellular movement of vesicles and organelles. Many antibodies against tubulins have been described, most of them against the C-terminal portion, which is exposed at the outside of the microtubules. By generating a novel set of monoclonal antibodies against the cytoskeleton of Trypanosoma cruzi, a flagellate protozoan that causes Chagas’ disease, we selected a clone (mAb 3G4) that recognizes β-tubulin. The epitope for mAb 3G4 was mapped by pepscan to a highly conserved sequence motif found between α-helices 11 and 12 of the C-terminus of β-tubulin in eukaryotes. It labels vesicular structures in both T. cruzi and mammalian cells, colocalizing respectively with a major cysteine protease (Cruzipain) and lysosome associated protein (LAMP2) respectively, but it does not label regular microtubules on these cellular models. We propose that the epitope recognized by mAb 3G4 is exposed only in a form of tubulin associated with endosomes.
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Keywords: Microtubules; Trypanosoma cruzi; Tubulin; chromosomes; endosome; epitope; lysosome; monoclonal antibodies; monoclonal antibody; parasite; post-translational modifications

Document Type: Research Article

Affiliations: Universidade Federal de Sao Paulo, Rua Pedro de Toledo 669 6ยบ Andar, 04039-032 Sao Paulo, SP, Brazil.

Publication date: 01 June 2012

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  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.
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