Journal Article

Targeting of proConA to the Plant Vacuole depends on its Nine Amino-acid C-terminal Propeptide

Saint-Jore-Dupas Claude, Gilbert Marie-Agnès, Ramis Catalina, Paris Nadine, Kiefer-Meyer Marie-Christine, Neuhaus Jean-Marc, Faye Loïc and Gomord Véronique

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 10, pages 1603-1612
Published in print October 2005 | ISSN: 0032-0781
Published online October 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci176
Targeting of proConA to the Plant Vacuole depends on its Nine Amino-acid C-terminal Propeptide

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  • Biochemistry
  • Molecular and Cell Biology
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Concanavalin A (ConA) is a well characterized and extensively used lectin accumulated in the protein bodies of jack bean cotyledons. ConA is synthesized as an inactive precursor proConA. The maturation of inactive proConA into biologically active ConA is a complex process including the removal of an internal glycopeptide and a C-terminal propeptide (CTPP), followed by a head-to-tail ligation of the two largest polypeptides. The cDNA encoding proConA was cloned and expressed in tobacco BY-2 cells. ProConA was slowly transported to the vacuole where its maturation into ConA was similar to that in jack bean cotyledons, apart from an incomplete final ligation. To investigate the role of the nine amino acid CTPP, a truncated form lacking the propeptide (proConAΔ9) was expressed in BY-2 cells. In contrast to proConA, proConAΔ9 was rapidly chased out of the endoplasmic reticulum (ER) and secreted into the culture medium. The CTPP was then fused to the C-terminal end of a secreted form of green fluorescent protein (secGFP). When expressed in tobacco BY-2 cells and leaf protoplasts, the chimaeric protein was located in the vacuole whereas secGFP was located in the culture medium and in the vacuole. Altogether, our results show we have isolated a new C-terminal vacuolar sorting determinant.

Keywords: BY-2 cells; Concanavalin A; GFP; Targeting; Vacuolar sorting determinant; Vacuole; 3D, three-dimensional; CE, cell extract; ConA, concanavalin A; CTPP, C-terminal propeptide; ctVSD, C-terminal VSD; ER, endoplasmic reticulum; EM, extracellular medium; GFP, green fluorescent protein; GFPChit, secGFP fused to chitinase vacuolar sorting signal; GFPConA, secGFP fused to ConA sorting signal; Mi, microsomal fraction; PEG, polyethylene glycol; PSV, protein storage vacuole; psVSD, physical structure VSD; S, soluble fraction; secGFP, secreted GFP; ssVSD, sequence specific VSD; VSD, vacuolar sorting determinant

Journal Article.  7362 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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