Compensatory rebalancing of rice prolamins by production of recombinant prolamin/bioactive peptide fusion proteins within ER-derived protein bodies.

Abstract

KEY MESSAGE:

Bioactive peptide was produced by fusion to rice prolamins in transgenic rice seeds. Their accumulation levels were affected by their deposition sites and by compensatory rebalancing between prolamins within PB-Is. Peptide immunotherapy using analogue peptide ligands (APLs) is one of promising treatments against autoimmune diseases. Use of seed storage protein as a fusion carrier is reasonable strategy for production of such small size bioactive peptides. In this study, to examine the efficacy of various rice prolamins deposited in ER-derived protein bodies (PB-Is), the APL12 from the Glucose-6-phosphate isomerase (GPI325-339) was expressed by fusion to four types of representative prolamins under the control of the individual native promoters. When the 14 and 16 kDa Cys-rich prolamins, which were localized in middle layer of PB-Is, were used for production of the APL12, they highly accumulated in transgenic rice seeds (~ 200 µg/grain). By contrast, fusion to the 10 and 13 kDa prolamins, which were localized in the core and outermost layer of PB-Is, resulted in lower levels of accumulation (~ 40 µg/grain). These results suggest that accumulation levels were highly affected by their deposition sites. Next, when different prolamin/APL12 fusion proteins were co-expressed to increase accumulation levels, they could not be increased so much as their expected additive levels. High accumulation of one type prolamin/APL12 led to reduction of other type(s) prolamin/APL12 to maintain the limited amounts of prolamins that can be deposited in PB-Is. Moreover, suppression of endogenous seed proteins by RNA interference also did not significantly enhance the accumulation levels of prolamin/APL12. These findings suggest that there may be compensatory rebalancing mechanism that controls the accumulation levels of prolamins deposited within PB-Is.

• Authors: Fumio Takaiwa, Lijun Yang, Yuhya Wakasa, Kenjiro Ozawa
• Journal: Plant cell reports (Plant Cell Rep) Vol. 37 Issue 2 Pg. 209-223 (Feb 2018) ISSN: 1432-203X [Electronic] Germany
• PMID: 29075848 (Publication Type: Journal Article)

Chemical References

• Peptides
• Plant Proteins
• Prolamins
• Recombinant Fusion Proteins

Topics

• Endosperm (genetics, metabolism)
• Gene Expression Regulation, Plant
• Immunoblotting
• Microscopy, Confocal
• Oryza (genetics, metabolism)
• Peptides (genetics, metabolism)
• Plant Proteins (genetics, metabolism)
• Plants, Genetically Modified
• Prolamins (genetics, metabolism)
• Recombinant Fusion Proteins (genetics, metabolism)
• Seeds (genetics, metabolism)