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Reconstitution of phospholipase A-dependent Golgi membrane tubules

Methods in Molecular Biology, ISSN: 1064-3745, Vol: 1496, Page: 75-90
2016
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Book Chapter Description

The Golgi complex is the Grand Central Station of intracellular membrane trafficking in the secretory and endocytic pathways. Anterograde and retrograde export of cargo from the Golgi complex involves a complex interplay between the formation of coated vesicles and membrane tubules, although much less is known about tubule-mediated trafficking. Recent advances using in vitro assays have identified several cytoplasmic phospholipase A(PLA) enzymes that are required for the biogenesis of membrane tubules and their roles in the functional organization of the Golgi complex. In this chapter we describe methods for the cell-free reconstitution of PLA-dependent Golgi membrane tubule formation. These methods should facilitate the identification of other proteins that regulate this process.

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