IBBR publication #1649

Chemical secretory pathway modulation in plant protoplasts

De Marchis F, Pompa A, Bellucci M

In: “Unconventional protein secretion” (Pompa A, De Marchis F eds). Springer, New York (USA), pp. 67-79. [ISBN: 978-1-4939-3804-9] (2016)
doi: 10.1007/978-1-4939-3804-9_4
URL: http://link.springer.com/protocol/10.1007/978-1-4939-3804-9_4

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.