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, 58 (2), 329-36

Purification of a Novel Class of Coated Vesicles Mediating Biosynthetic Protein Transport Through the Golgi Stack

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Purification of a Novel Class of Coated Vesicles Mediating Biosynthetic Protein Transport Through the Golgi Stack

V Malhotra et al. Cell.

Abstract

We describe a scheme for the purification of the nonclathrin-coated vesicles that mediate transport of proteins between Golgi cisternae and probably from ER to Golgi. These "Golgi-derived coated vesicles" accumulate when Golgi membranes are incubated with ATP and cytosol in the presence of GTP gamma S, a compound that blocks vesicle fusion. The coated vesicles dissociate from the Golgi cisternae in high salt and can then be purified by employing differential and density gradient centrifugation. Golgi-derived coated vesicles have a putative polypeptide composition that is distinct from both cytosol and Golgi membranes, as well as from that of clathrin-coated vesicles.

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