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
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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