Eukaryotic protein production in designed storage organelles

M. Torrent, B. Llompart, S. Lasserre-Ramassamy, I. Llop-Tous, M. Bastida, P. Marzabal, Ann Westerholm-Parvinen, Markku Saloheimo, P. B. Heifetz (Corresponding Author), M. D. Ludevid (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

89 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number5
JournalBMC Biology
Volume7
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

protein bodies
Organelles
organelles
recombinant proteins
protein
Recombinant Proteins
Proteins
proteins
endoplasmic reticulum
Endoplasmic Reticulum
tobacco
Seed
biopharmaceuticals
Trichoderma reesei
Tobacco
zein
insects
seed storage proteins
insulating materials
Insects

Keywords

  • endoplasmic reticulum
  • vacuole
  • protein bodies
  • organelles
  • eukaryotic cell

Cite this

Torrent, M., Llompart, B., Lasserre-Ramassamy, S., Llop-Tous, I., Bastida, M., Marzabal, P., ... Ludevid, M. D. (2009). Eukaryotic protein production in designed storage organelles. BMC Biology, 7, [5]. https://doi.org/10.1186/1741-7007-7-5
Torrent, M. ; Llompart, B. ; Lasserre-Ramassamy, S. ; Llop-Tous, I. ; Bastida, M. ; Marzabal, P. ; Westerholm-Parvinen, Ann ; Saloheimo, Markku ; Heifetz, P. B. ; Ludevid, M. D. / Eukaryotic protein production in designed storage organelles. In: BMC Biology. 2009 ; Vol. 7.
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title = "Eukaryotic protein production in designed storage organelles",
abstract = "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.",
keywords = "endoplasmic reticulum, vacuole, protein bodies, organelles, eukaryotic cell",
author = "M. Torrent and B. Llompart and S. Lasserre-Ramassamy and I. Llop-Tous and M. Bastida and P. Marzabal and Ann Westerholm-Parvinen and Markku Saloheimo and Heifetz, {P. B.} and Ludevid, {M. D.}",
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Torrent, M, Llompart, B, Lasserre-Ramassamy, S, Llop-Tous, I, Bastida, M, Marzabal, P, Westerholm-Parvinen, A, Saloheimo, M, Heifetz, PB & Ludevid, MD 2009, 'Eukaryotic protein production in designed storage organelles', BMC Biology, vol. 7, 5. https://doi.org/10.1186/1741-7007-7-5

Eukaryotic protein production in designed storage organelles. / Torrent, M.; Llompart, B.; Lasserre-Ramassamy, S.; Llop-Tous, I.; Bastida, M.; Marzabal, P.; Westerholm-Parvinen, Ann; Saloheimo, Markku; Heifetz, P. B. (Corresponding Author); Ludevid, M. D. (Corresponding Author).

In: BMC Biology, Vol. 7, 5, 2009.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Eukaryotic protein production in designed storage organelles

AU - Torrent, M.

AU - Llompart, B.

AU - Lasserre-Ramassamy, S.

AU - Llop-Tous, I.

AU - Bastida, M.

AU - Marzabal, P.

AU - Westerholm-Parvinen, Ann

AU - Saloheimo, Markku

AU - Heifetz, P. B.

AU - Ludevid, M. D.

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

KW - endoplasmic reticulum

KW - vacuole

KW - protein bodies

KW - organelles

KW - eukaryotic cell

U2 - 10.1186/1741-7007-7-5

DO - 10.1186/1741-7007-7-5

M3 - Article

VL - 7

JO - BMC Biology

JF - BMC Biology

SN - 1741-7007

M1 - 5

ER -

Torrent M, Llompart B, Lasserre-Ramassamy S, Llop-Tous I, Bastida M, Marzabal P et al. Eukaryotic protein production in designed storage organelles. BMC Biology. 2009;7. 5. https://doi.org/10.1186/1741-7007-7-5