The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence
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....
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2014-12-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fbioe.2014.00067/full |
| _version_ | 1818303027455459328 |
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| author | Anna eHofbauer Jenny ePeters Elsa eArcalis Thomas eRademacher Johannes eLampel Francois eEudes Alessandro eVitale Eva eStoger |
| author_facet | Anna eHofbauer Jenny ePeters Elsa eArcalis Thomas eRademacher Johannes eLampel Francois eEudes Alessandro eVitale Eva eStoger |
| author_sort | Anna eHofbauer |
| collection | DOAJ |
| description | 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 kD 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. |
| first_indexed | 2024-12-13T05:48:16Z |
| format | Article |
| id | doaj.art-4d19635277d246d994ff4611d4e66cf2 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-13T05:48:16Z |
| publishDate | 2014-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-4d19635277d246d994ff4611d4e66cf22022-12-21T23:57:37ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852014-12-01210.3389/fbioe.2014.00067122213The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequenceAnna eHofbauer0Jenny ePeters1Elsa eArcalis2Thomas eRademacher3Johannes eLampel4Francois eEudes5Alessandro eVitale6Eva eStoger7University of Natural Resources and Life SCiencesUniversity of Natural Resources and Life SCiencesUniversity of Natural Resources and Life SCiencesRWTH AachenUniversity of Natural Resources and Life SCiencesAgriculture and Agri-Food CanadaCNRUniversity of Natural Resources and Life SCiencesNaturally-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 kD 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.http://journal.frontiersin.org/Journal/10.3389/fbioe.2014.00067/fullMolecular Farmingsubcellular targetingintermembrane spaceProtein Bodiesplastid importrecombinent protein |
| spellingShingle | Anna eHofbauer Jenny ePeters Elsa eArcalis Thomas eRademacher Johannes eLampel Francois eEudes Alessandro eVitale Eva eStoger The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence Frontiers in Bioengineering and Biotechnology Molecular Farming subcellular targeting intermembrane space Protein Bodies plastid import recombinent protein |
| title | The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence |
| title_full | The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence |
| title_fullStr | The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence |
| title_full_unstemmed | The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence |
| title_short | The induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid-targeting signal in the 27 kD γ-zein sequence |
| title_sort | induction of recombinant protein bodies in different subcellular compartments reveals a cryptic plastid targeting signal in the 27 kd γ zein sequence |
| topic | Molecular Farming subcellular targeting intermembrane space Protein Bodies plastid import recombinent protein |
| url | http://journal.frontiersin.org/Journal/10.3389/fbioe.2014.00067/full |
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