Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m

Human cardiotrophin 1 (CT1), a cytokine with excellent therapeutic potential, was previously expressed in tobacco chloroplasts. However, the growth conditions required to reach the highest expression levels resulted in an impairment of its bioactivity. In the present study, we have examined new stra...

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Autores: Ancín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059ac, Fernández-San Millán, A. (Alicia)|||/items/fcc31688-5b60-4171-a12a-c2beca8a01c3, Santamaría, E. (Eva)|||/items/7060efc6-8410-47f8-a7c4-51845be6a4a3, Larraya, L. (Luis)|||/items/fadc066a-439a-45d0-907c-106539bae9d6, Veramendi, J. (Jon)|||/items/ad8670eb-b12e-454d-aa6c-a6e036724156, Farran, I. (Inmaculada)|||/items/0f398365-05ac-4596-87bf-df4d2c6735bc, Sanz-Barrio, R. (Ruth)|||/items/f4daa266-5dc7-4757-8682-3ccdec8f2b86
Tipo de recurso: artículo
Fecha de publicación:2020
País:España
Institución:Universidad de Navarra
Repositorio:Dadun. Depósito Académico Digital de la Universidad de Navarra
Idioma:inglés
OAI Identifier:oai:dadun.unav.edu:10171/66320
Acceso en línea:https://hdl.handle.net/10171/66320
Access Level:acceso abierto
Palabra clave:Cardiotrophin-1
Thioredoxin
Plastid transformation
Bioactivity
Tobacco
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spelling Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin mAncín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059acFernández-San Millán, A. (Alicia)|||/items/fcc31688-5b60-4171-a12a-c2beca8a01c3Santamaría, E. (Eva)|||/items/7060efc6-8410-47f8-a7c4-51845be6a4a3Larraya, L. (Luis)|||/items/fadc066a-439a-45d0-907c-106539bae9d6Veramendi, J. (Jon)|||/items/ad8670eb-b12e-454d-aa6c-a6e036724156Farran, I. (Inmaculada)|||/items/0f398365-05ac-4596-87bf-df4d2c6735bcSanz-Barrio, R. (Ruth)|||/items/f4daa266-5dc7-4757-8682-3ccdec8f2b86Cardiotrophin-1ThioredoxinPlastid transformationBioactivityTobaccoHuman cardiotrophin 1 (CT1), a cytokine with excellent therapeutic potential, was previously expressed in tobacco chloroplasts. However, the growth conditions required to reach the highest expression levels resulted in an impairment of its bioactivity. In the present study, we have examined new strategies to modulate the expression of this recombinant protein in chloroplasts so as to enhance its production and bioactivity. In particular, we assessed the effect of both the fusion and co-expression of Trx m with CT1 on the production of a functional CT1 by using plastid transformation. Our data revealed that the Trx m fusion strategy was useful to increase the expression levels of CT1 inside the chloroplasts, although CT1 bioactivity was significantly impaired, and this was likely due to steric hindrance between both proteins. By contrast, the expression of functional CT1 was increased when co-expressed with Trx m, because we demonstrated that recombinant CT1 was functionally active during an in vitro signaling assay. While Trx m/CT1 co-expression did not increase the amount of CT1 in young leaves, our results revealed an increase in CT1 protein stability as the leaves aged in this genotype, which also improved the recombinant protein’s overall production. This strategy might be useful to produce other functional biopharmaceuticals in chloroplasts.Dadun. Depósito Académico Digital Universidad de Navarra20232023-05-2320202020-01-0120202020-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10171/66320reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/663202026-06-21T12:47:57Z
dc.title.none.fl_str_mv Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
title Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
spellingShingle Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
Ancín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059ac
Cardiotrophin-1
Thioredoxin
Plastid transformation
Bioactivity
Tobacco
title_short Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
title_full Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
title_fullStr Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
title_full_unstemmed Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
title_sort Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m
dc.creator.none.fl_str_mv Ancín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059ac
Fernández-San Millán, A. (Alicia)|||/items/fcc31688-5b60-4171-a12a-c2beca8a01c3
Santamaría, E. (Eva)|||/items/7060efc6-8410-47f8-a7c4-51845be6a4a3
Larraya, L. (Luis)|||/items/fadc066a-439a-45d0-907c-106539bae9d6
Veramendi, J. (Jon)|||/items/ad8670eb-b12e-454d-aa6c-a6e036724156
Farran, I. (Inmaculada)|||/items/0f398365-05ac-4596-87bf-df4d2c6735bc
Sanz-Barrio, R. (Ruth)|||/items/f4daa266-5dc7-4757-8682-3ccdec8f2b86
author Ancín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059ac
author_facet Ancín, M. (María)|||/items/145c9698-fe55-4af8-aa59-cc429c6059ac
Fernández-San Millán, A. (Alicia)|||/items/fcc31688-5b60-4171-a12a-c2beca8a01c3
Santamaría, E. (Eva)|||/items/7060efc6-8410-47f8-a7c4-51845be6a4a3
Larraya, L. (Luis)|||/items/fadc066a-439a-45d0-907c-106539bae9d6
Veramendi, J. (Jon)|||/items/ad8670eb-b12e-454d-aa6c-a6e036724156
Farran, I. (Inmaculada)|||/items/0f398365-05ac-4596-87bf-df4d2c6735bc
Sanz-Barrio, R. (Ruth)|||/items/f4daa266-5dc7-4757-8682-3ccdec8f2b86
author_role author
author2 Fernández-San Millán, A. (Alicia)|||/items/fcc31688-5b60-4171-a12a-c2beca8a01c3
Santamaría, E. (Eva)|||/items/7060efc6-8410-47f8-a7c4-51845be6a4a3
Larraya, L. (Luis)|||/items/fadc066a-439a-45d0-907c-106539bae9d6
Veramendi, J. (Jon)|||/items/ad8670eb-b12e-454d-aa6c-a6e036724156
Farran, I. (Inmaculada)|||/items/0f398365-05ac-4596-87bf-df4d2c6735bc
Sanz-Barrio, R. (Ruth)|||/items/f4daa266-5dc7-4757-8682-3ccdec8f2b86
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Dadun. Depósito Académico Digital Universidad de Navarra
dc.subject.none.fl_str_mv Cardiotrophin-1
Thioredoxin
Plastid transformation
Bioactivity
Tobacco
topic Cardiotrophin-1
Thioredoxin
Plastid transformation
Bioactivity
Tobacco
description Human cardiotrophin 1 (CT1), a cytokine with excellent therapeutic potential, was previously expressed in tobacco chloroplasts. However, the growth conditions required to reach the highest expression levels resulted in an impairment of its bioactivity. In the present study, we have examined new strategies to modulate the expression of this recombinant protein in chloroplasts so as to enhance its production and bioactivity. In particular, we assessed the effect of both the fusion and co-expression of Trx m with CT1 on the production of a functional CT1 by using plastid transformation. Our data revealed that the Trx m fusion strategy was useful to increase the expression levels of CT1 inside the chloroplasts, although CT1 bioactivity was significantly impaired, and this was likely due to steric hindrance between both proteins. By contrast, the expression of functional CT1 was increased when co-expressed with Trx m, because we demonstrated that recombinant CT1 was functionally active during an in vitro signaling assay. While Trx m/CT1 co-expression did not increase the amount of CT1 in young leaves, our results revealed an increase in CT1 protein stability as the leaves aged in this genotype, which also improved the recombinant protein’s overall production. This strategy might be useful to produce other functional biopharmaceuticals in chloroplasts.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-01-01
2020
2020-01-01
2023
2023-05-23
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/10171/66320
url https://hdl.handle.net/10171/66320
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra
instname:Universidad de Navarra
instname_str Universidad de Navarra
reponame_str Dadun. Depósito Académico Digital de la Universidad de Navarra
collection Dadun. Depósito Académico Digital de la Universidad de Navarra
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repository.mail.fl_str_mv
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score 15.300719

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