期刊
BIOTECHNOLOGY JOURNAL
卷 10, 期 9, 页码 1478-1486出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201500056
关键词
Influenza M2; Ion channel expression; Intracellular pH; Recombinant hemagglutinin; Transient expression
资金
- Natural Science and Engineering Research Council (NSERC) of Canada
- NSERC
- Fonds de Recherche Quebec/Nature et Technologies
- Medicago inc.
Eukaryotic expression systems are used for the production of complex secreted proteins. How ever, recombinant proteins face considerable biochemical challenges along the secretory pathway, including proteolysis and pH variation between organelles. As the use of synthetic biology matures into solutions for protein production, various host-cell engineering approaches are being developed to ameliorate host-cell factors that can limit recombinant protein quality and yield. We report the potential of the influenza M2 ion channel as a novel tool to neutralize the pH in acidic subcellular compartments. Using transient expression in the plant host Nicotiana benthamiana, we show that ion channel expression can significantly raise pH in the Golgi apparatus and that this can have a strong stabilizing effect on a fusion protein separated by an acid-susceptible linker peptide. We exemplify the utility of this effect in recombinant protein production using influenza hemagglutinin subtypes differentially stable at low pH; the expression of hemagglutinins prone to conformational change in mildly acidic conditions is considerably enhanced by M2 co-expression. The co-expression of a heterologous ion channel to stabilize acid-labile proteins and peptides represents a novel approach to increasing the yield and quality of secreted recombinant proteins in plants and, possibly, in other eukaryotic expression hosts.
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