Eukaryotic protein production in designed storage organelles
Autor: | Margarita Torrent, Sabine Lasserre-Ramassamy, M. Dolors Ludevid, Peter B Heifetz, Ann Westerholm-Parvinen, Miriam Bastida, Markku Saloheimo, Pau Marzabal, Blanca Llompart, Immaculada Llop-Tous |
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Přispěvatelé: | European Commission, Ministerio de Ciencia y Tecnología (España), Generalitat de Catalunya |
Rok vydání: | 2009 |
Předmět: |
Physiology
Zein Sf9 CHO Cells Plant Science Biology Zea mays General Biochemistry Genetics and Molecular Biology eukaryotic cell Cell Line law.invention Zera Cricetulus Structural Biology law Cricetinae Tobacco Organelle Protein biosynthesis Animals Storage protein lcsh:QH301-705.5 Ecology Evolution Behavior and Systematics Plant Proteins Trichoderma chemistry.chemical_classification Agricultural and Biological Sciences(all) vacuole Biochemistry Genetics and Molecular Biology(all) Endoplasmic reticulum Chinese hamster ovary cell Cell Biology biology.organism_classification Molecular biology Recombinant Proteins Cell biology endoplasmic reticulum Eukaryotic Cells protein bodies organelles lcsh:Biology (General) Gene Expression Regulation chemistry Recombinant DNA General Agricultural and Biological Sciences Research Article Developmental Biology Biotechnology |
Zdroj: | Dipòsit Digital de Documents de la UAB Universitat Autònoma de Barcelona Recercat. Dipósit de la Recerca de Catalunya instname BMC Biology Digital.CSIC. Repositorio Institucional del CSIC Recercat: Dipósit de la Recerca de Catalunya Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya) BMC Biology, Vol 7, Iss 1, p 5 (2009) Torrent, M, Llompart, B, Lasserre-Ramassamy, S, Llop-Tous, I, Bastida, M, Marzabal, P, Westerholm-Parvinen, A, Saloheimo, M, Heifetz, P B & Ludevid, M D 2009, ' Eukaryotic protein production in designed storage organelles ', BMC Biology, vol. 7, 5 . https://doi.org/10.1186/1741-7007-7-5 |
Popis: | [Background] Protein bodies (PBs) are natural endoplasmic reticulum (ER) or vacuole plant-derived organelles that stably accumulate large amounts of storage proteins in seeds. The proline-rich N-terminal domain derived from the maize storage protein γ zein (Zera) is sufficient to induce PBs in non-seed tissues of Arabidopsis and tobacco. This Zera property opens up new routes for high-level accumulation of recombinant proteins by fusion of Zera with proteins of interest. In this work we extend the advantageous properties of plant seed PBs to recombinant protein production in useful non-plant eukaryotic hosts including cultured fungal, mammalian and insect cells. [Results] Various Zera fusions with fluorescent and therapeutic proteins accumulate in induced PB-like organelles in all eukaryotic systems tested: tobacco leaves, Trichoderma reesei, several mammalian cultured cells and Sf9 insect cells. This accumulation in membranous organelles insulates both recombinant protein and host from undesirable activities of either. Recombinant protein encapsulation in these PBs facilitates stable accumulation of proteins in a protected sub-cellular compartment which results in an enhancement of protein production without affecting the viability and development of stably transformed hosts. The induced PBs also retain the high-density properties of native seed PBs which facilitate the recovery and purification of the recombinant proteins they contain. [Conclusion] The Zera sequence provides an efficient and universal means to produce recombinant proteins by accumulation in ER-derived organelles. The remarkable cross-kingdom conservation of PB formation and their biophysical properties should have broad application in the manufacture of non-secreted recombinant proteins and suggests the existence of universal ER pathways for protein insulation. SLR was supported by a postdoctoral fellowship from the Marie Curie Program and PM was the recipient of a RED postdoctoral grant (Generalitat de Catalunya). This work was supported by grants to MDL from MCYT-FEDER (BIO 2004-03202), the Generalitat de Calalunya (CeRBa and 2005 SGR00182) and ERA Biotech, S.A. |
Databáze: | OpenAIRE |
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