title: VAMP8-3xHA Uptake Assay in HeLa Cells.
abstract: Transmembrane proteins are rarely exclusively localized to a specific vesicle or an organelle. Most transmembrane proteins undergo complicated trafficking routes. Thus, transmembrane proteins are under constant flux, and at steady state, found on a variety of vesicles or organelles. This characteristic makes the study of their trafficking routes complex, since at any given moment, different molecules are often being trafficked in opposing directions. Pulse-chase experiments can temporally track a specific pool of a transmembrane protein of interest, allowing for the kinetic description of its trafficking route. This type of technique has been used extensively to follow a large array of plasma membrane localized proteins (Dirilet al., 2006; Jeanet al., 2010). Here, we describe a method that allows the study of VAMP8 trafficking from the plasma membrane to endolysosomal compartments. This method was used to describe a role forMTMR13andRAB21in the regulation of VAMP8 trafficking to endolysosomes (Jeanet al., 2015).
authors.affiliation: Département d'anatomie et de biologie cellulaire, Faculté de Médecine et des Sciences de la Santé. Université de Sherbrooke, Sherbrooke, Canada.
authors.forename: Steve
authors.initials: S
authors.lastname: Jean
authors.affiliation: Division of Biological Sciences, University of California, San Diego, USA.
authors.forename: Amy A
authors.initials: AA
authors.lastname: Kiger