Subcellular targeting of human interleukin-10 in plants.
The utility of plants for the production of a wide range of recombinant proteins is now clearly established. However, the challenge remains to produce these proteins at sufficient concentrations for extraction to be economically feasible. In this paper, we have investigated the ability of plant cells to accumulate the human interleukin-10 (IL-10) protein targeted to chloroplasts and mitochondria. We found that IL-10 accumulates in chloroplasts only if a 6 x His tag is added at the C-terminus of the protein. The hexapeptide may provide protection from degradation. Conversely, the IL-10 protein does not accumulate in mitochondria. Analysis of the chloroplast-targeted IL-10 protein revealed only monomeric IL-10 and limited biological activity in in vitro cell assays.
Menassa
Rima
R
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, Ont., Canada N5V 4T3.
Kennette
Wendy
W
Nguyen
Vi
V
Rymerson
Robert
R
Jevnikar
Anthony
A
Brandle
Jim
J
D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism*; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D016753:Interleukin-10