Sorting of phaseolin to the vacuole is saturable and requires a short C-terminal peptide.
Phaseolin, one of the major legume proteins for human nutrition, is a trimeric glycoprotein of the 7S class that accumulates in the protein storage vacuoles of common bean. Phaseolin is cotranslationally introduced into the lumen of the endoplasmic reticulum; from there, it is transported through the Golgi complex to the storage vacuoles. Phaseolin is also transported to the vacuole in vegetative tissues of transgenic plants. By transient and permanent expression in tobacco leaf cells, we show here that vacuolar sorting of phaseolin is saturable and that saturation leads to Golgi-mediated secretion from the cell. A mutated phaseolin, in which the four C-terminal residues (Ala, Phe, Val, and Tyr) were deleted, efficiently formed trimers but was secreted entirely outside of the cells in transgenic tobacco leaves, indicating that the deleted sequence contains information necessary for interactions with the saturable vacuolar sorting machinery. In the apoplast, the secreted phaseolin remained intact; this is similar to what occurs to wild-type phaseolin in bean storage vacuoles, whereas in vegetative vacuoles of transgenic plants, the storage protein is fragmented.
Frigerio
L
L
Istituto Biosintesi Vegetali, Consiglio Nazionale delle Ricerche, via Bassini 15, 20133 Milan, Italy.
de Virgilio
M
M
Prada
A
A
Faoro
F
F
Vitale
A
A
D000595:Amino Acid Sequence; D007887:Fabaceae / Q000502:physiology*; D006056:Golgi Apparatus / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009747:Nutritional Physiological Phenomena; D018515:Plant Leaves / Q000378:metabolism / Q000648:ultrastructure; D010940:Plant Proteins; D030821:Plants, Genetically Modified; D010946:Plants, Medicinal; D010947:Plants, Toxic; D011523:Protoplasts / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D014026:Nicotiana / Q000378:metabolism; D014118:Toxins, Biological / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure
D010940:Plant Proteins; D011994:Recombinant Proteins; D014118:Toxins, Biological; C048826:phaseolin protein, Phaseolus vulgaris