Chemical Secretory Pathway Modulation in Plant Protoplasts.
The classical Golgi pathway is not the only mechanism for vacuolar protein transport in plants because alternative transport mechanisms have been described. The existence of these alternative pathways can be demonstrated using several chemicals and here we describe the use of brefeldin A (BFA), endo-β-N-acetylglucosaminidase H (Endo-H), and tunicamycin, on isolated tobacco leaf protoplasts. Two main methods are illustrated in this chapter, protoplast pulse-chase followed by protein immunoprecipitation, and protoplast immunofluorescence.
Institute of Biosciences and Bioresources, Research Division of Perugia, National Research Council, 06128, Perugia, Italy.
Francesca
F
De Marchis
Institute of Biosciences and Bioresources, Research Division of Perugia, National Research Council, 06128, Perugia, Italy.
Andrea
A
Pompa
Institute of Biosciences and Bioresources, Research Division of Perugia, National Research Council, 06128, Perugia, Italy. michele.bellucci@ibbr.cnr.it.
Michele
M
Bellucci
D001692:Biological Transport; D006026:Glycoside Hydrolases / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D047468:Immunoprecipitation; D018515:Plant Leaves / Q000378:metabolism; D010944:Plants / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D021381:Protein Transport; D011523:Protoplasts / Q000378:metabolism*; D055571:Secretory Pathway; D014026:Nicotiana / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D011134:Polysaccharides; D006026:Glycoside Hydrolases
BFA; Endo-H; Golgi-mediated protein traffic; Pulse-chase; Tobacco; Tunicamycin