Delivery of a secreted soluble protein to the vacuole via a membrane anchor.
To further understand how membrane proteins are sorted in the secretory system, we devised a strategy that involves the expression of a membrane-anchored yeast invertase in transgenic plants. The construct consisted of a signal peptide followed by the coding region of yeast invertase and the transmembrane domain and cytoplasmic tail of calnexin. The substitution of a lysine near the C terminus of calnexin with a glutamic acid residue ensured progression through the secretory system rather than retention in or return to the endoplasmic reticulum. In the transformed plants, invertase activity and a 70-kD cross-reacting protein were found in the vacuoles. This yeast invertase had plant-specific complex glycans, indicating that transport to the vacuole was mediated by the Golgi apparatus. The microsomal fraction contained a membrane-anchored 90-kD cross-reacting polypeptide, but was devoid of invertase activity. Our results indicate that this membrane-anchored protein proceeds in the secretory system beyond the point where soluble proteins are sorted for secretion, and is detached from its membrane anchor either just before or just after delivery to the vacuole.
Barrieu
F
F
Department of Biology, University of California at San Diego, La Jolla, California 92093-0116, USA.
Chrispeels
M J
MJ
D000595:Amino Acid Sequence; D001692:Biological Transport; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D037281:Calnexin; D002451:Cell Compartmentation; D003429:Cross Reactions; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008861:Microsomes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012995:Solubility; D014026:Nicotiana; D014617:Vacuoles / Q000201:enzymology / Q000378:metabolism*; D043324:beta-Fructofuranosidase
D002135:Calcium-Binding Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D037281:Calnexin; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase