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Hofbauer A, Peters J, Arcalis E, et al. The Induction of Recombinant Protein Bodies in Different Subcellular Compartments Reveals a Cryptic Plastid-Targeting Signal in the 27-kDa γ-Zein Sequence. Front Bioeng Biotechnol. 2014;2:67doi: 10.3389/fbioe.2014.00067.
Hofbauer, A., Peters, J., Arcalis, E., Rademacher, T., Lampel, J., Eudes, F., Vitale, A., & Stoger, E. (2014). The Induction of Recombinant Protein Bodies in Different Subcellular Compartments Reveals a Cryptic Plastid-Targeting Signal in the 27-kDa γ-Zein Sequence. Frontiers in bioengineering and biotechnology, 267. https://doi.org/10.3389/fbioe.2014.00067
Hofbauer, Anna, et al. "The Induction of Recombinant Protein Bodies in Different Subcellular Compartments Reveals a Cryptic Plastid-Targeting Signal in the 27-kDa γ-Zein Sequence." Frontiers in bioengineering and biotechnology vol. 2 (2014): 67. doi: https://doi.org/10.3389/fbioe.2014.00067
Hofbauer A, Peters J, Arcalis E, Rademacher T, Lampel J, Eudes F, Vitale A, Stoger E. The Induction of Recombinant Protein Bodies in Different Subcellular Compartments Reveals a Cryptic Plastid-Targeting Signal in the 27-kDa γ-Zein Sequence. Front Bioeng Biotechnol. 2014 Dec 11;2:67. doi: 10.3389/fbioe.2014.00067. eCollection 2014. PMID: 25566533; PMCID: PMC4263181.
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Front Bioeng Biotechnol. 2014 Dec 11;2:67. doi: 10.3389/fbioe.2014.00067. eCollection 2014.

The Induction of Recombinant Protein Bodies in Different Subcellular Compartments Reveals a Cryptic Plastid-Targeting Signal in the 27-kDa γ-Zein Sequence.

Frontiers in bioengineering and biotechnology

Anna Hofbauer, Jenny Peters, Elsa Arcalis, Thomas Rademacher, Johannes Lampel, François Eudes, Alessandro Vitale, Eva Stoger

Affiliations

  1. Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences , Vienna , Austria.
  2. Institute of Molecular Biotechnology, RWTH Aachen University , Aachen , Germany.
  3. Agriculture and Agri-Food Canada , Lethbridge, AB , Canada.
  4. Institute of Agricultural Biology and Biotechnology, National Research Council (CNR) , Milan , Italy.

PMID: 25566533 PMCID: PMC4263181 DOI: 10.3389/fbioe.2014.00067

Abstract

Naturally occurring storage proteins such as zeins are used as fusion partners for recombinant proteins because they induce the formation of ectopic storage organelles known as protein bodies (PBs) where the proteins are stabilized by intermolecular interactions and the formation of disulfide bonds. Endogenous PBs are derived from the endoplasmic reticulum (ER). Here, we have used different targeting sequences to determine whether ectopic PBs composed of the N-terminal portion of mature 27 kDa γ-zein added to a fluorescent protein could be induced to form elsewhere in the cell. The addition of a transit peptide for targeting to plastids causes PB formation in the stroma, whereas in the absence of any added targeting sequence PBs were typically associated with the plastid envelope, revealing the presence of a cryptic plastid-targeting signal within the γ-zein cysteine-rich domain. The subcellular localization of the PBs influences their morphology and the solubility of the stored recombinant fusion protein. Our results indicate that the biogenesis and budding of PBs does not require ER-specific factors and therefore, confirm that γ-zein is a versatile fusion partner for recombinant proteins offering unique opportunities for the accumulation and bioencapsulation of recombinant proteins in different subcellular compartments.

Keywords: intermembrane space; molecular farming; plastid import; protein bodies; recombinant protein; subcellular targeting

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