Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major
Main Author: | |
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Publication Date: | 2021 |
Other Authors: | , , , , , , , |
Format: | Article |
Language: | eng |
Source: | Repositório Institucional da UNESP |
Download full: | http://dx.doi.org/10.1016/j.biochi.2020.12.017 http://hdl.handle.net/11449/209226 |
Summary: | Heat shock proteins (Hsps) are involved in several important aspects of the cell proteostasis. Hsp90 interacts with at least a tenth of the cell proteome helping a large number of proteins to fold correctly. Hsp90 function is modulated by several co-chaperones having TPR (tetratricopeptide repeat) domains that allow for interaction with the C-terminal MEEVD motif of the chaperone. Another important chaperone, Hsp70, has a C-terminal EEVD motif that binds to TPR. Leishmania is a protozoan that causes leishmaniasis, a neglected disease in humans and other animals. There is still no effective treatment for leishmaniasis, however the study of structure and function of the proteins of the parasite may generate potential targets for future therapeutic intervention studies. In this work, the genome of Leishmania major was searched for a novel TPR-domain gene, which is conserved in Leishmania. The recombinant protein, LmTPR, was produced in pure and folded state and was characterized by biophysical tools as a monomer with an elongated conformation. Studies in Leishmania major were also preformed to complement these in vitro experiments. Splice Leader RNA-seq analysis and Western blot indicated that the protein was expressed in all developmental stages of the parasite. Binding assays confirmed that both Hsp90 and Hsp70 bind specifically to LmTPR. Finally, sequence and structural predictions indicated a C terminal region as a RPAP3 domain. Altogether, this study identified a novel TPR-domain co-chaperone of Hsp90 that is conserved and expressed in all developmental stages of Leishmania major. (C) 2021 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. |
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Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania majorTPR-Domain proteinLeishmania majorHsp90 co-chaperonesProtein foldingProtein structure and functionHeat shock proteins (Hsps) are involved in several important aspects of the cell proteostasis. Hsp90 interacts with at least a tenth of the cell proteome helping a large number of proteins to fold correctly. Hsp90 function is modulated by several co-chaperones having TPR (tetratricopeptide repeat) domains that allow for interaction with the C-terminal MEEVD motif of the chaperone. Another important chaperone, Hsp70, has a C-terminal EEVD motif that binds to TPR. Leishmania is a protozoan that causes leishmaniasis, a neglected disease in humans and other animals. There is still no effective treatment for leishmaniasis, however the study of structure and function of the proteins of the parasite may generate potential targets for future therapeutic intervention studies. In this work, the genome of Leishmania major was searched for a novel TPR-domain gene, which is conserved in Leishmania. The recombinant protein, LmTPR, was produced in pure and folded state and was characterized by biophysical tools as a monomer with an elongated conformation. Studies in Leishmania major were also preformed to complement these in vitro experiments. Splice Leader RNA-seq analysis and Western blot indicated that the protein was expressed in all developmental stages of the parasite. Binding assays confirmed that both Hsp90 and Hsp70 bind specifically to LmTPR. Finally, sequence and structural predictions indicated a C terminal region as a RPAP3 domain. Altogether, this study identified a novel TPR-domain co-chaperone of Hsp90 that is conserved and expressed in all developmental stages of Leishmania major. (C) 2021 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CIHRUniv Campinas UNICAMP, Inst Chem, BR-13083970 Campinas, SP, BrazilNatl Inst Sci & Technol Struct Biol & Bioimage IN, Rio De Janeiro, RJ, BrazilSao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, BrazilUniv Sao Paulo, Sao Carlos Inst Chem, Sao Carlos, SP, BrazilUniv Toronto, Dept Biochem, Toronto, ON M5G 1M1, CanadaUniv Toronto, Dept Chem, Toronto, ON M5S 3H6, CanadaSao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, BrazilFAPESP: 2012/50161-8FAPESP: 2017/26131-5FAPESP: 2018/04375-2FAPESP: 2014/25967-4FAPESP: 2019/11496-3CAPES: 99999.004913/2015-09CIHR: PJT-173491Elsevier B.V.Universidade Estadual de Campinas (UNICAMP)Natl Inst Sci & Technol Struct Biol & Bioimage INUniversidade Estadual Paulista (Unesp)Universidade de São Paulo (USP)Univ TorontoAraujo, Sara A.Martins, Gustavo H.Quel, Natalia G.Aragao, Annelize Z. B.Morea, Edna G. O. [UNESP]Borges, Julio C.Houry, Walid A.Cano, Maria I. N. [UNESP]Ramos, Carlos H.2021-06-25T11:52:11Z2021-06-25T11:52:11Z2021-03-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article51-60http://dx.doi.org/10.1016/j.biochi.2020.12.017Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 182, p. 51-60, 2021.0300-9084http://hdl.handle.net/11449/20922610.1016/j.biochi.2020.12.017WOS:000620773000005Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiochimieinfo:eu-repo/semantics/openAccess2021-10-23T19:23:38Zoai:repositorio.unesp.br:11449/209226Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T19:23:38Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
title |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
spellingShingle |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major Araujo, Sara A. TPR-Domain protein Leishmania major Hsp90 co-chaperones Protein folding Protein structure and function |
title_short |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
title_full |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
title_fullStr |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
title_full_unstemmed |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
title_sort |
Purification and characterization of a novel and conserved TPR-domain protein that binds both Hsp90 and Hsp70 and is expressed in all developmental stages of Leishmania major |
author |
Araujo, Sara A. |
author_facet |
Araujo, Sara A. Martins, Gustavo H. Quel, Natalia G. Aragao, Annelize Z. B. Morea, Edna G. O. [UNESP] Borges, Julio C. Houry, Walid A. Cano, Maria I. N. [UNESP] Ramos, Carlos H. |
author_role |
author |
author2 |
Martins, Gustavo H. Quel, Natalia G. Aragao, Annelize Z. B. Morea, Edna G. O. [UNESP] Borges, Julio C. Houry, Walid A. Cano, Maria I. N. [UNESP] Ramos, Carlos H. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual de Campinas (UNICAMP) Natl Inst Sci & Technol Struct Biol & Bioimage IN Universidade Estadual Paulista (Unesp) Universidade de São Paulo (USP) Univ Toronto |
dc.contributor.author.fl_str_mv |
Araujo, Sara A. Martins, Gustavo H. Quel, Natalia G. Aragao, Annelize Z. B. Morea, Edna G. O. [UNESP] Borges, Julio C. Houry, Walid A. Cano, Maria I. N. [UNESP] Ramos, Carlos H. |
dc.subject.por.fl_str_mv |
TPR-Domain protein Leishmania major Hsp90 co-chaperones Protein folding Protein structure and function |
topic |
TPR-Domain protein Leishmania major Hsp90 co-chaperones Protein folding Protein structure and function |
description |
Heat shock proteins (Hsps) are involved in several important aspects of the cell proteostasis. Hsp90 interacts with at least a tenth of the cell proteome helping a large number of proteins to fold correctly. Hsp90 function is modulated by several co-chaperones having TPR (tetratricopeptide repeat) domains that allow for interaction with the C-terminal MEEVD motif of the chaperone. Another important chaperone, Hsp70, has a C-terminal EEVD motif that binds to TPR. Leishmania is a protozoan that causes leishmaniasis, a neglected disease in humans and other animals. There is still no effective treatment for leishmaniasis, however the study of structure and function of the proteins of the parasite may generate potential targets for future therapeutic intervention studies. In this work, the genome of Leishmania major was searched for a novel TPR-domain gene, which is conserved in Leishmania. The recombinant protein, LmTPR, was produced in pure and folded state and was characterized by biophysical tools as a monomer with an elongated conformation. Studies in Leishmania major were also preformed to complement these in vitro experiments. Splice Leader RNA-seq analysis and Western blot indicated that the protein was expressed in all developmental stages of the parasite. Binding assays confirmed that both Hsp90 and Hsp70 bind specifically to LmTPR. Finally, sequence and structural predictions indicated a C terminal region as a RPAP3 domain. Altogether, this study identified a novel TPR-domain co-chaperone of Hsp90 that is conserved and expressed in all developmental stages of Leishmania major. (C) 2021 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-25T11:52:11Z 2021-06-25T11:52:11Z 2021-03-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.biochi.2020.12.017 Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 182, p. 51-60, 2021. 0300-9084 http://hdl.handle.net/11449/209226 10.1016/j.biochi.2020.12.017 WOS:000620773000005 |
url |
http://dx.doi.org/10.1016/j.biochi.2020.12.017 http://hdl.handle.net/11449/209226 |
identifier_str_mv |
Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 182, p. 51-60, 2021. 0300-9084 10.1016/j.biochi.2020.12.017 WOS:000620773000005 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biochimie |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
51-60 |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1797790233667829760 |