Unexpected deposition patterns of recombinant proteins in post-endoplasmic reticulum compartments of wheat endosperm.
Protein transport within cereal endosperm cells is complicated by the abundance of endoplasmic reticulum (ER)-derived and vacuolar protein bodies. For wheat storage proteins, two major transport routes run from the ER to the vacuole, one bypassing and one passing through the Golgi. Proteins traveling along each route converge at the vacuole and form aggregates. To determine the impact of this trafficking system on the fate of recombinant proteins expressed in wheat endosperm, we used confocal and electron microscopy to investigate the fate of three recombinant proteins containing different targeting information. KDEL-tagged recombinant human serum albumin, which is retrieved to the ER lumen in leaf cells, was deposited in prolamin aggregates within the vacuole of endosperm cells, most likely following the bulk of endogenous glutenins. Recombinant fungal phytase, a glycoprotein designed for secretion, was delivered to the same compartment, with no trace of the molecule in the apoplast. Glycan analysis revealed that this protein had passed through the Golgi. The localization of human serum albumin and phytase was compared to that of recombinant legumin, which contains structural targeting information directing it to the vacuole. Uniquely, legumin accumulated in the globulin inclusion bodies at the periphery of the prolamin bodies, suggesting a different mode of transport and/or aggregation. Our results demonstrate that recombinant proteins are deposited in an unexpected pattern within wheat endosperm cells, probably because of the unique storage properties of this tissue. Our data also confirm that recombinant proteins are invaluable tools for the analysis of protein trafficking in cereals.
Institute for Molecular Biotechnology , Rheinische-Westfälische Technische Hochschule Aachen, 52074 Aachen, Germany.
Elsa
E
Arcalis
Sylvain
S
Marcel
Friedrich
F
Altmann
Daniel
D
Kolarich
Georgia
G
Drakakaki
Rainer
R
Fischer
Paul
P
Christou
Eva
E
Stoger
D010832:6-Phytase / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000378:metabolism; D055315:Prolamins; D021382:Protein Sorting Signals; D021381:Protein Transport; D011994:Recombinant Proteins / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism* / Q000648:ultrastructure; D012709:Serum Albumin / Q000378:metabolism; D014908:Triticum / Q000378:metabolism* / Q000648:ultrastructure; D000097793:Legumins
D010832:6-Phytase; D010940:Plant Proteins; D055315:Prolamins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D012709:Serum Albumin; D000097793:Legumins