Release of putative exocytic transport vesicles from perforated MDCK cells.
Mechanically perforated MDCK cells were used to study membrane transport between the trans-Golgi network and the apical and basolateral plasma membrane domains in vitro. Three membrane transport markers--an apical protein (fowl plague virus haemagglutinin), a basolateral protein (vesicular stomatitis virus G protein), and a lipid marker destined for both domains (C6-NBD-sphingomyelin)--were each accumulated in the trans-Golgi by a 20 degrees C block of transport and their behaviour monitored following cell perforation and incubation at 37 degrees C. In the presence of ATP and in the absence of calcium ions a considerable fraction of the transport markers were released from the perforated cells in sealed membrane vesicles. Control experiments showed that the vesicles were not generated by non-specific vesiculation of the Golgi complex or the plasma membrane. The vesicles had well defined sedimentation properties and the orientation expected of transport vesicles derived from the trans-Golgi network.
Bennett
M K
MK
European Molecular Biology Laboratory, Heidelberg, FRG.
Wandinger-Ness
A
A
Simons
K
K
D000818:Animals; D001693:Biological Transport, Active; D015415:Biomarkers; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D005089:Exocytosis; D006056:Golgi Apparatus / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism
D015415:Biomarkers