Molecular characterization and intracellular distribution of the alpha 5 subunit of Trypanosoma cruzi 20S proteasome Journal Article
| Authors: | Gutiérrez, B.; Osorio, L.; Motta, M. C. M.; Huima-Byron, T.; Erdjument-Bromage, H.; Munoz, C.; Sagua, H.; Mortara, R. A.; Echeverría, A.; Araya, J. E.; Gonzalez, J. |
| Article Title: | Molecular characterization and intracellular distribution of the alpha 5 subunit of Trypanosoma cruzi 20S proteasome |
| Abstract: | Three different monoclonal antibodies were produced against Trypanosona cruzi proteasomes. These antibodies were shown to react with a single 27-kDa band on immunoblots of purified proteasomes. Using a 7E5 monoclonal antibody (IgG1) that recognized the α5 subunit of protozoan protease we have studied the intracellular distribution of the T. cruzi 20S proteasome. Contrary to all cell types described to date, T. cruzi 20S proteasome was found not only in the cytoplasm and nucleus but also in the kinetoplast. As revealed by confocal microscopy, the reactivity of monoclonal antibody 7E5 was highly specific for protozoan proteasome because the antibody recognized only the proteasomes from parasites and not those from the mammalian host in T. cruzi infected cells. These findings were confirmed by immunoblots or immunoprecipitations, followed by chymotrypsin-like activity detection in kinetoplasts isolated by differential centrifugation and sucrose density gradients. Proteasome 20S was present in all T. cruzi stages and only slight differences in terms of relative abundance were found. The potential role of the proteasome in kinetoplast remodeling remains to be determined. © 2009. |
| Keywords: | immunohistochemistry; controlled study; unclassified drug; nonhuman; animal cell; mouse; mammalia; animals; proteasome; proteasome endopeptidase complex; confocal microscopy; microscopy, confocal; immunoreactivity; cell type; fluorescent antibody technique; antibodies, monoclonal; enzyme analysis; molecular recognition; immunoprecipitation; immunoblotting; cellular distribution; cytoplasm; antibody specificity; cell nucleus; antibodies; protein subunits; enzyme subunit; intracellular localization; trypanosoma cruzi; 20s proteasome; chymotrypsin; immunoglobulin g1 antibody; sucrose; density gradient; enzyme localization; enzyme purification; host cell; kinetoplast; centrifugation, density gradient; life cycle stages; subcellular fractions; protozoa |
| Journal Title: | Parasitology International |
| Volume: | 58 |
| Issue: | 4 |
| ISSN: | 1383-5769 |
| Publisher: | Elsevier Ireland Ltd. |
| Date Published: | 2009-12-01 |
| Start Page: | 367 |
| End Page: | 374 |
| Language: | English |
| DOI: | 10.1016/j.parint.2009.07.012 |
| PUBMED: | 19666140 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Export Date: 30 November 2010" - "CODEN: PAINF" - "Source: Scopus" |
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