title: A recombinant multimeric immunoglobulin expressed in rice shows assembly-dependent subcellular localization in endosperm cells.
abstract: To investigate the role of subunit assembly in the intracellular deposition of multimeric recombinant proteins, we expressed a partially humanized secretory immunoglobulin in rice endosperm cells and determined the subcellular locations of the assembled protein and its individual components. Transgenic rice plants expressing either individual subunits or all the subunits of the antibody were generated by particle bombardment, and protein localization was determined by immunoelectron microscopy. Assembly of the antibody was confirmed by immunoassay and coimmunoprecipitation. Immunolocalization experiments showed no evidence for secretion of the antibody or any of its components to the apoplast. Rather, the nonassembled light chain, heavy chain and secretory component accumulated predominantly within endoplasmic reticulum-derived protein bodies, while the assembled antibody, with antigen-binding function, accumulated specifically in protein storage vacuoles. These results show that the destination of a complex recombinant protein within the plant cell is influenced by its state of assembly.
authors.affiliation: John Innes Centre, Colney Lane, Norwich NR4 7UH, Norfolk, UK.
authors.forename: Liz
authors.initials: L
authors.lastname: Nicholson
authors.forename: Pablo
authors.initials: P
authors.lastname: Gonzalez-Melendi
authors.forename: Craig
authors.initials: C
authors.lastname: van Dolleweerd
authors.forename: Heidi
authors.initials: H
authors.lastname: Tuck
authors.forename: Yolande
authors.initials: Y
authors.lastname: Perrin
authors.forename: Julian K-C
authors.initials: JK
authors.lastname: Ma
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.forename: Paul
authors.initials: P
authors.lastname: Christou
authors.forename: Eva
authors.initials: E
authors.lastname: Stoger