Eukaryotic protein production in designed storage organelles
Torrent, Margarita (Institut de Recerca i Tecnologia Agroalimentàries)
Llompart, Blanca (ERA Biotech. S.A. Parc Científic de Barcelona)
Lasserre-Ramassamy, Sabine (Institut de Recerca i Tecnologia Agroalimentàries)
Llop-Tous, Immaculada (Institut de Recerca i Tecnologia Agroalimentàries)
Bastida, Miriam (ERA Biotech. S.A. Parc Científic de Barcelona)
Marzabal, Pau (ERA Biotech. S.A. Parc Científic de Barcelona)
Westerholm-Pavinen, Ann (VTT Technical Research Centre)
Saloheimo, Markku (VTT Technical Research)
Heifetz, Peter B. (ERA Biotech. S.A. Parc Científic de Barcelona)
Ludevid, M. Dolors (Institut de Recerca i Tecnologia Agroalimentàries)

Fecha:	2009
Resumen:	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.
Ayudas:	Ministerio de Ciencia y Tecnología BIO2004-03202 Agència de Gestió d'Ajuts Universitaris i de Recerca 2005/SGR-00182
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Publicado en:	BMC biology, Vol. 7 (January 2009) , art. 5, ISSN 1741-7007

DOI: 10.1186/1741-7007-7-5
PMID: 19175916

14 p, 4.6 MB

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Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias > CRAG (Centro de Investigación en Agrigenómica)
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