Endogenous glycosphingolipids move to the cell surface at a rate consistent with bulk flow estimates.
The bulk flow model of intracellular trafficking predicts that forward transport from the ER through the Golgi to the plasma membrane proceeds by default without a special signal being required (Wieland, F.T., Gleason, M. L., Serafini, T. A., and Rothman, J. E. (1987) Cell 50, 289-300). We tested a crucial prediction of this model, which is that the endogenous lipid components of the transport vesicles would reach the plasma membrane at the rapid rate of bulk flow. The rate at which endogenous glycosphingolipids moved from the ER through the Golgi to the plasma membrane was determined in Chinese hamster ovary cells using metabolic labeling with tritiated palmitate and oxidation of cell surface ganglioside NeuAc alpha 2----3Gal beta 1----4Glc beta 1----4Cer (GM3) with periodate. Whereas radioactive precursor became incorporated into ceramide and glucosyl ceramide without a detectable lag, synthesis of labeled lactosyl ceramide and ganglioside GM3 did not begin until 5-6 min and 11-12 min, respectively, after addition of labeled precursor. Labeled GM3 reached the plasma membrane 5-6 min following its synthesis. Overall, approximately 18 min transpired from the time that the ceramide precursor was synthesized in the ER until labeled GM3 reached the plasma membrane. These results indicate that lipid transport vesicles move rapidly to the plasma membrane at a rate consistent with bulk flow estimates.
Young
W W
WW
Department of Biological and Biophysical Sciences, University of Louisville, Kentucky 40292.
Lutz
M S
MS
Blackburn
W A
WA
D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005679:G(M3) Ganglioside / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism
D005679:G(M3) Ganglioside; D006028:Glycosphingolipids; D010169:Palmitic Acids; D019308:Palmitic Acid