title: Plastoglobules: a new address for targeting recombinant proteins in the chloroplast.
abstract: BACKGROUND
  The potential of transgenic plants for cost-effective production of pharmaceutical molecules is now becoming apparent. Plants have the advantage over established fermentation systems (bacterial, yeast or animal cell cultures) to circumvent the risk of pathogen contamination, to be amenable to large scaling up and to necessitate only established farming procedures. Chloroplasts have proven a useful cellular compartment for protein accumulation owing to their large size and number, as well as the possibility for organellar transformation. They therefore represent the targeting destination of choice for recombinant proteins in leaf crops such as tobacco. Extraction and purification of recombinant proteins from leaf material contribute to a large extent to the production costs. Developing new strategies facilitating these processes is therefore necessary.
  
  
  RESULTS
  Here, we evaluated plastoglobule lipoprotein particles as a new subchloroplastic destination for recombinant proteins. The yellow fluorescent protein as a trackable cargo was targeted to plastoglobules when fused to plastoglobulin 34 (PGL34) as the carrier. Similar to adipocyte differentiation related protein (ADRP) in animal cells, most of the protein sequence of PGL34 was necessary for targeting to lipid bodies. The recombinant protein was efficiently enriched in plastoglobules isolated by simple flotation centrifugation. The viability of plants overproducing the recombinant protein was not affected, indicating that plastoglobule targeting did not significantly impair photosynthesis or sugar metabolism.
  
  
  CONCLUSIONS
  Our data identify plastoglobules as a new targeting destination for recombinant protein in leaf crops. The wide-spread presence of plastoglobules and plastoglobulins in crop species promises applications comparable to those of transgenic oilbody-oleosin technology in molecular farming.
authors.affiliation: Institute of Botany, University of Neuchâtel, rue Emile Argand 11, CP158, CH-2009 Neuchâtel, Switzerland. pvidi@purdue.edu <pvidi@purdue.edu>
authors.forename: Pierre-Alexandre
authors.initials: PA
authors.lastname: Vidi
authors.forename: Felix
authors.initials: F
authors.lastname: Kessler
authors.forename: Claire
authors.initials: C
authors.lastname: Bréhélin
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry; D002241:Carbohydrates; D002736:Chloroplasts / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D005821:Genetic Techniques; D008055:Lipids / Q000737:chemistry; D008164:Luminescent Proteins / Q000737:chemistry; D010788:Photosynthesis; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry*; D010944:Plants / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry*; D014026:Nicotiana / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D002241:Carbohydrates; D008055:Lipids; D008164:Luminescent Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins; C054562:yellow fluorescent protein, Bacteria; D004247:DNA