Separation of early steps in endocytic membrane transport

Goot, F. Gisou van der

doi:10.1002/elps.1150181426

Cited by 19 publications

(16 citation statements)

References 22 publications

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“…As a control, the distribution of the toxin was analyzed in cells that had been treated with ASSP at 4°C. The surface‐bound toxin was in a non‐reduced state and was highly enriched in fraction 2, as expected since this fraction contains plasma membrane rafts (van der Goot, 1997).…”

Section: Resultssupporting

confidence: 73%

Differential sorting and fate of endocytosed GPI-anchored proteins

Fivaz¹,

Vilbois²,

Thurnheer³

et al. 2002

The EMBO Journal

204

208

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In this paper, we studied the fate of endocytosed glycosylphosphatidyl inositol anchored proteins (GPIAPs) in mammalian cells, using aerolysin, a bacterial toxin that binds to the GPI anchor, as a probe. We ®nd that GPI-APs are transported down the endocytic pathway to reducing late endosomes in BHK cells, using biochemical, morphological and functional approaches. We also ®nd that this transport correlates with the association to raft-like membranes and thus that lipid rafts are present in late endosomes (in addition to the Golgi and the plasma membrane). In marked contrast, endocytosed GPI-APs reach the recycling endosome in CHO cells and this transport correlates with a decreased raft association. GPI-APs are, however, diverted from the recycling endosome and routed to late endosomes in CHO cells, when their raft association is increased by clustering seven or less GPI-APs with an aerolysin mutant. We conclude that the different endocytic routes followed by GPI-APs in different cell types depend on the residence time of GPI-APs in lipid rafts, and hence that raft partitioning regulates GPI-APs sorting in the endocytic pathway.

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Section: Resultssupporting

confidence: 73%

Differential sorting and fate of endocytosed GPI-anchored proteins

Fivaz¹,

Vilbois²,

Thurnheer³

et al. 2002

The EMBO Journal

204

208

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“…As previously, late endosomes were obtained from Baby Hamster Kidney cells, since a well-established subcellular fractionation protocol to purify late endosomes is available for this cell line [25], [32]. We have previously shown using surface biotinylation of proteins that this fraction does not contain detectable amounts of plasma membrane [33]. Moreover this late endosomal fraction is not contaminated by Golgi, endoplasmic reticulum, early endosomal or caveolar membranes, as shown by western blotting using marker proteins [34].…”

Section: Resultsmentioning

confidence: 86%

Regulation of the V-ATPase along the Endocytic Pathway Occurs through Reversible Subunit Association and Membrane Localization

et al. 2008

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The lumen of endosomal organelles becomes increasingly acidic when going from the cell surface to lysosomes. Luminal pH thereby regulates important processes such as the release of internalized ligands from their receptor or the activation of lysosomal enzymes. The main player in endosomal acidification is the vacuolar ATPase (V-ATPase), a multi-subunit transmembrane complex that pumps protons from the cytoplasm to the lumen of organelles, or to the outside of the cell. The active V-ATPase is composed of two multi-subunit domains, the transmembrane V0 and the cytoplasmic V1. Here we found that the ratio of membrane associated V1/Vo varies along the endocytic pathway, the relative abundance of V1 being higher on late endosomes than on early endosomes, providing an explanation for the higher acidity of late endosomes. We also found that all membrane-bound V-ATPase subunits were associated with detergent resistant membranes (DRM) isolated from late endosomes, raising the possibility that association with lipid-raft like domains also plays a role in regulating the activity of the proton pump. In support of this, we found that treatment of cells with U18666A, a drug that leads to the accumulation of cholesterol in late endosomes, affected acidification of late endosome. Altogether our findings indicate that the activity of the vATPase in the endocytic pathway is regulated both by reversible association/dissociation and the interaction with specific lipid environments.

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“…marker of plasma membrane and early endosomes (13). Annexin II was highly concentrated in plasma membrane compared with whole cell lysate and was weakly present in the cytosolic S 1 fraction from unstimulated cells (Fig.…”

Section: Resultsmentioning

confidence: 87%

“…The uncoated endocytotic vesicles are contained in the light membrane fraction that is included in the V 2 pellet (12). We determined also that the ␣-synuclein signal increase in the cytoplasmic S 1 fraction was matched by an increase in the annexin II signal, a marker for early endosomes (13) (Fig. 3).…”

Section: Resultsmentioning

confidence: 89%

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Muscarinic Receptor Stimulation Induces Translocation of an α-Synuclein Oligomer from Plasma Membrane to a Light Vesicle Fraction in Cytoplasm

Leng¹,

Chase²,

Bennett³

2001

Journal of Biological Chemistry

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The close correspondence between the distribution of brain ␣-synuclein and that of muscarinic M 1 and M 3 receptors suggests a role for this protein in cholinergic transmission. We thus examined the effect of muscarinic stimulation on ␣-synuclein in SH-SY5Y, a human dopaminergic cell line that expresses this protein. Under basal conditions, ␣-synuclein was detected in all subcellular compartments isolated as follows: plasma membrane, cytoplasm, nucleus, and two vesicle fractions. The lipid fractions contained only a 45-kDa ␣-synuclein oligomer, whereas the cytoplasmic and nuclear fractions contained both the oligomer and the monomer. This finding suggests ␣-synuclein exists physiologically as a lipid-bound oligomer and a soluble monomer. Muscarinic stimulation by carbachol reduced the ␣-synuclein oligomer in plasma membrane over a 30-min period, with a concomitant increase of both the oligomer and the monomer in the cytoplasmic fraction. The oligomer was associated with a light vesicle fraction in cytoplasm that contains uncoated endocytotic vesicles. The carbachol-induced alteration of ␣-synuclein was blocked by atropine. Translocation of the ␣-synuclein oligomer in response to carbachol stimulation corresponds closely with the time course of ligandstimulated muscarinic receptor endocytosis. The data suggest that the muscarine receptor stimulated release of the ␣-synuclein oligomer from plasma membrane, and its subsequent association with the endocytotic vesicle fraction may have a role in muscarine receptor endocytosis. We propose that its function may be a transient release of membrane-bound phospholipase D 2 from ␣-synuclein inhibition, thus allowing this lipase to participate in muscarinic receptor endocytosis.The neuronal protein ␣-synuclein, recently implicated in the pathogenesis of Parkinson's disease (1-3), has a brain distribution closely matching that of muscarinic M 1 and M 3 receptors and the muscarine receptor-linked enzymes phospholipase C and protein kinase C (4). The proximity of ␣-synuclein to brain muscarinic receptors and related enzymes suggests that this protein could contribute to cholinergic function. To evaluate this possibility, we investigated the effect of muscarinic receptor stimulation by the cholinergic agonist carbachol on the subcellular distribution of endogenous ␣-synuclein in the human neuroblastoma cell line SH-SY5Y. This dopaminergic cell line shares many properties with dopamine neurons of substantia nigra pars compacta, the major locus of neurodegeneration in Parkinson's disease, including the expression of M 1 and M 3 receptors (5). We now report that carbachol induces a translocation of a 45-kDa ␣-synuclein oligomer from plasma membrane to a light vesicle fraction in cytoplasm, with a time course and subcellular localization matching that of muscarinic receptors during carbachol-stimulated endocytosis. Previously, synucleins have been demonstrated to be endogenous inhibitors of phospholipase D 2 (PLD 2 ) 1 (6). Furthermore, activation of PLD isoforms, including PLD 2 , ...

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Separation of early steps in endocytic membrane transport

Cited by 19 publications

References 22 publications

Differential sorting and fate of endocytosed GPI-anchored proteins

Differential sorting and fate of endocytosed GPI-anchored proteins

Regulation of the V-ATPase along the Endocytic Pathway Occurs through Reversible Subunit Association and Membrane Localization

Muscarinic Receptor Stimulation Induces Translocation of an α-Synuclein Oligomer from Plasma Membrane to a Light Vesicle Fraction in Cytoplasm

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