Abstract
Publisher Summary This chapter describe a method that utilizes semiintact Chinese hamster ovary (CHO) cells to reconstitute vesicular traffic from the trans-Golgi region to the plasma membrane. The assay utilizes a bacterial cytolysin, streptolysin O (SL-O), to introduce large aqueous pores selectively into the plasma membrane. The general application of SL-O to permeabilize cells is reviewed. As this method produces minimal damage to the cell surface, it is suitable for studying various transport pathways between the trans-Golgi and the plasma membrane. This approach is successfully utilized by the group to study intracellular transport in CHO cells. The trans-Golgi is a key compartment involved in tracking among the cellular organelles. Newly synthesized proteins to be released into the extracellular space, incorporated into the plasma membrane, or localized to lysosomes all traverse the same compartments until they reach the trans-Golgi. At the trans-Golgi network, these proteins are sorted from one another and packaged into unique classes of vesicles that are targeted to distinct post-Golgi destinations.
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