Unintended molecular interactions in transgenic plants expressing clinically useful proteins: the case of bovine aprotinin traveling the potato leaf cell secretory pathway.
We assessed the impact of subcellular targeting on the heterologous expression of a clinically useful protease inhibitor, bovine aprotinin, in leaves of potato, Solanum tuberosum. Transgenic potato lines targeting aprotinin to the cytosol, the ER or the apoplast were first generated, and then assessed for their ability to accumulate the recombinant protein. On-chip detection and quantitation of aprotinin variants by SELDI TOF MS showed the inhibitor to be absent in the cytosol, but present under different forms in the ER and the apoplast. No visible phenotypic effects of aprotinin were observed for the transgenic lines, but aprotinin retention in the ER was associated with a significant decrease of leaf soluble protein content. A 2-D gel assessment of control and transgenic lines revealed a possible link between this altered protein content and the down-regulation of proteins implicated in protein synthesis and maturation. These observations, supported by complementary 2-DE analyses with potato lines targeting aprotinin to the apoplast, suggest an aprotinin-mediated feedback in planta negatively altering protein anabolism. From a practical viewpoint, these data illustrate the importance of taking into account not only the characteristics of recombinant proteins expressed in heterologous environments, but also their possible effects on protein accumulation in the host plant factory.
Badri
M Amine
MA
Département de phytologie, CRH/INAF, Université Laval, Québec, Canada.
Rivard
Daniel
D
Coenen
Karine
K
Michaud
Dominique
D
D000818:Animals; D007611:Aprotinin / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D055571:Secretory Pathway / Q000235:genetics / Q000502:physiology*; D011198:Solanum tuberosum / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D007611:Aprotinin