The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone.
BACKGROUND: Trypanosoma cruzi has a single flagellum attached to the cell body by a network of specialized cytoskeletal and membranous connections called the flagellum attachment zone. Previously, we isolated a DNA fragment (clone H49) which encodes tandemly arranged repeats of 68 amino acids associ...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3214072?pdf=render |
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| author | Alexandra Galetović Renata T Souza Marcia R M Santos Esteban M Cordero Izabela M D Bastos Jaime M Santana Jeronimo C Ruiz Fabio M Lima Marjorie M Marini Renato A Mortara José Franco da Silveira |
| author_facet | Alexandra Galetović Renata T Souza Marcia R M Santos Esteban M Cordero Izabela M D Bastos Jaime M Santana Jeronimo C Ruiz Fabio M Lima Marjorie M Marini Renato A Mortara José Franco da Silveira |
| author_sort | Alexandra Galetović |
| collection | DOAJ |
| description | BACKGROUND: Trypanosoma cruzi has a single flagellum attached to the cell body by a network of specialized cytoskeletal and membranous connections called the flagellum attachment zone. Previously, we isolated a DNA fragment (clone H49) which encodes tandemly arranged repeats of 68 amino acids associated with a high molecular weight cytoskeletal protein. In the current study, the genomic complexity of H49 and its relationships to the T. cruzi calpain-like cysteine peptidase family, comprising active calpains and calpain-like proteins, is addressed. Immunofluorescence analysis and biochemical fractionation were used to demonstrate the cellular location of H49 proteins. METHODS AND FINDINGS: All of H49 repeats are associated with calpain-like sequences. Sequence analysis demonstrated that this protein, now termed H49/calpain, consists of an amino-terminal catalytic cysteine protease domain II, followed by a large region of 68-amino acid repeats tandemly arranged and a carboxy-terminal segment carrying the protease domains II and III. The H49/calpains can be classified as calpain-like proteins as the cysteine protease catalytic triad has been partially conserved in these proteins. The H49/calpains repeats share less than 60% identity with other calpain-like proteins in Leishmania and T. brucei, and there is no immunological cross reaction among them. It is suggested that the expansion of H49/calpain repeats only occurred in T. cruzi after separation of a T. cruzi ancestor from other trypanosomatid lineages. Immunofluorescence and immunoblotting experiments demonstrated that H49/calpain is located along the flagellum attachment zone adjacent to the cell body. CONCLUSIONS: H49/calpain contains large central region composed of 68-amino acid repeats tandemly arranged. They can be classified as calpain-like proteins as the cysteine protease catalytic triad is partially conserved in these proteins. H49/calpains could have a structural role, namely that of ensuring that the cell body remains attached to the flagellum by connecting the subpellicular microtubule array to it. |
| first_indexed | 2024-12-11T20:02:21Z |
| format | Article |
| id | doaj.art-531fcc1d2e374685925b71e5252e916f |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T20:02:21Z |
| publishDate | 2011-01-01 |
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| series | PLoS ONE |
| spelling | doaj.art-531fcc1d2e374685925b71e5252e916f2022-12-22T00:52:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01611e2763410.1371/journal.pone.0027634The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone.Alexandra GaletovićRenata T SouzaMarcia R M SantosEsteban M CorderoIzabela M D BastosJaime M SantanaJeronimo C RuizFabio M LimaMarjorie M MariniRenato A MortaraJosé Franco da SilveiraBACKGROUND: Trypanosoma cruzi has a single flagellum attached to the cell body by a network of specialized cytoskeletal and membranous connections called the flagellum attachment zone. Previously, we isolated a DNA fragment (clone H49) which encodes tandemly arranged repeats of 68 amino acids associated with a high molecular weight cytoskeletal protein. In the current study, the genomic complexity of H49 and its relationships to the T. cruzi calpain-like cysteine peptidase family, comprising active calpains and calpain-like proteins, is addressed. Immunofluorescence analysis and biochemical fractionation were used to demonstrate the cellular location of H49 proteins. METHODS AND FINDINGS: All of H49 repeats are associated with calpain-like sequences. Sequence analysis demonstrated that this protein, now termed H49/calpain, consists of an amino-terminal catalytic cysteine protease domain II, followed by a large region of 68-amino acid repeats tandemly arranged and a carboxy-terminal segment carrying the protease domains II and III. The H49/calpains can be classified as calpain-like proteins as the cysteine protease catalytic triad has been partially conserved in these proteins. The H49/calpains repeats share less than 60% identity with other calpain-like proteins in Leishmania and T. brucei, and there is no immunological cross reaction among them. It is suggested that the expansion of H49/calpain repeats only occurred in T. cruzi after separation of a T. cruzi ancestor from other trypanosomatid lineages. Immunofluorescence and immunoblotting experiments demonstrated that H49/calpain is located along the flagellum attachment zone adjacent to the cell body. CONCLUSIONS: H49/calpain contains large central region composed of 68-amino acid repeats tandemly arranged. They can be classified as calpain-like proteins as the cysteine protease catalytic triad is partially conserved in these proteins. H49/calpains could have a structural role, namely that of ensuring that the cell body remains attached to the flagellum by connecting the subpellicular microtubule array to it.http://europepmc.org/articles/PMC3214072?pdf=render |
| spellingShingle | Alexandra Galetović Renata T Souza Marcia R M Santos Esteban M Cordero Izabela M D Bastos Jaime M Santana Jeronimo C Ruiz Fabio M Lima Marjorie M Marini Renato A Mortara José Franco da Silveira The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. PLoS ONE |
| title | The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. |
| title_full | The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. |
| title_fullStr | The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. |
| title_full_unstemmed | The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. |
| title_short | The repetitive cytoskeletal protein H49 of Trypanosoma cruzi is a calpain-like protein located at the flagellum attachment zone. |
| title_sort | repetitive cytoskeletal protein h49 of trypanosoma cruzi is a calpain like protein located at the flagellum attachment zone |
| url | http://europepmc.org/articles/PMC3214072?pdf=render |
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