Distinct transport vesicles mediate the delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells.
Open Access
- 1 September 1990
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 111 (3), 987-1000
- https://doi.org/10.1083/jcb.111.3.987
Abstract
Immunoisolation techniques have led to the purification of apical and basolateral transport vesicles that mediate the delivery of proteins from the trans-Golgi network to the two plasma membrane domains of MDCK cells. We showed previously that these transport vesicles can be formed and released in the presence of ATP from mechanically perforated cells (Bennett, M. K., A. Wandinger-Ness, and K. Simons, 1988. EMBO (Euro. Mol. Biol. Organ.) J. 7:4075-4085). Using virally infected cells, we have monitored the purification of the trans-Golgi derived vesicles by following influenza hemagglutinin or vesicular stomatitis virus (VSV) G protein as apical and basolateral markers, respectively. Equilibrium density gradient centrifugation revealed that hemagglutinin containing vesicles had a slightly lower density than those containing VSV-G protein, indicating that the two fractions were distinct. Antibodies directed against the cytoplasmically exposed domains of the viral spike glycoproteins permitted the resolution of apical and basolateral vesicle fractions. The immunoisolated vesicles contained a subset of the proteins present in the starting fraction. Many of the proteins were sialylated as expected for proteins existing the trans-Golgi network. The two populations of vesicles contained a number of proteins in common, as well as components which were enriched up to 38-fold in one fraction relative to the other. Among the unique components, a number of transmembrane proteins could be identified using Triton X-114 phase partitioning. This work provides evidence that two distinct classes of vesicles are responsible for apical and basolateral protein delivery. Common protein components are suggested to be involved in vesicle budding and fusion steps, while unique components may be required for specific recognition events such as those involved in protein sorting and vesicle targeting.Keywords
This publication has 63 references indexed in Scilit:
- Isolation of exocytic carrier vesicles from BHK cellsCell, 1989
- Vesicle fusion following receptor-mediated endocytosis requires a protein active in Golgi transportNature, 1989
- Dissection of a single round of vesicular transport: Sequential intermediates for intercisternal movement in the Golgi stackCell, 1989
- Role of an N-ethylmaleimide-sensitive transport component in promoting fusion of transport vesicles with cisternae of the Golgi stackCell, 1988
- A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeastCell, 1988
- Alteration of the cytoplasmic domain of the membrane-spanning glycoprotein p62 of Semliki Forest virus does not affect its polar distribution in established lines of Madin-Darby canine kidney cells.The Journal of cell biology, 1986
- The trans Golgi Network: Sorting at the Exit Site of the Golgi ComplexScience, 1986
- A new type of coated vesicular carrier that appears not to contain clathrin: Its possible role in protein transport within the Golgi stackCell, 1986
- Sequential intermediates in the pathway of intercompartmental transport in a cell-free systemCell, 1984
- Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylationCell, 1983