Ultrastructural localization of flotillin-1 to cholesterol-rich membrane microdomains, rafts, in rat brain tissue

Kokubo, Hideko; Helms, J. Bernd; Ohno‐Iwashita, Yoshiko; Shimada, Yukiko; Horikoshi, Y.; Yamaguchi, Hideyo

doi:10.1016/s0006-8993(02)04140-9

Cited by 58 publications

(55 citation statements)

References 26 publications

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“…The detergent resistance of the enlargeosome membrane opened now the possibility of a link to the other detergent-resistant organelles, in particular to caveolin-rich vesicles, caveosomes, and other known raft-rich vesicles trafficking between the TGN and the plasma membrane. Of the three raft markers investigated, however, caveolin1 was found to lack any colocalization with d/A, whereas Thy1 and also flotillin1, which is localized not in a single type of membrane but is distributed to various types (Morrow et al, 2002;Kokubo et al, 2003), exhibited only a low degree of colocalization. These results exclude the identification of enlargeosomes with organelles specific for the three markers.…”

Section: Discussionmentioning

confidence: 94%

Enlargeosome, an Exocytic Vesicle Resistant to Nonionic Detergents, Undergoes Endocytosis via a Nonacidic Route

Cocucci

Racchetti

Podini

et al. 2004

MBoC

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Section: Discussionmentioning

confidence: 94%

Enlargeosome, an Exocytic Vesicle Resistant to Nonionic Detergents, Undergoes Endocytosis via a Nonacidic Route

Cocucci

Racchetti

Podini

et al. 2004

MBoC

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“…Thus, sustained PKC stimulation resulted in sequestration of several recycling membrane proteins (CD59, transferrin receptor, and caveolin) but not molecules destined for the lysosomes (LDL, DiIC 16 (3)) or other membrane proteins, such as flotillin, a proposed lipid raft marker that has also been seen to accumulate in maturing phagosomes (38) and in lysosomes and Golgi (39). Thus, this newly recognized subset of PKC-influenced cargo appears to be composed of proteins and lipids that are known to be continuously recycled in the cell and that form at least a subset of recycling endosomes.…”

Section: Discussionmentioning

confidence: 98%

Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C

Idkowiak‐Baldys

Becker

Kitatani

et al. 2006

Journal of Biological Chemistry

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It has been previously shown that upon sustained stimulation (30 -60 min) with phorbol esters, protein kinase C (PKC) ␣ and ␤II become sequestered in a juxtanuclear region, the pericentrion. The activation of PKC also results in sequestration of transferrin, suggesting a role for PKC in regulating endocytosis and sequestration of recycling components. In this work we characterize the pericentrion as a PKC-dependent subset of the recycling compartment. We demonstrate that upon sustained stimulation of PKC, both protein (CD59, caveolin) and possibly also lipid (Bodipy-GM1) cargo become sequestered in a PKC-dependent manner. This sequestration displayed a strict temperature requirement and was inhibited below 32°C. Treatment of cells with phorbol myristate acetate for 60 min led to the formation of a distinct membrane structure. PKC sequestration and pericentrion formation were blocked by hypertonic sucrose as well as by potassium depletion (inhibitors of clathrin-dependent endocytosis) but not by nystatin or filipin, which inhibit clathrin-independent pathways. Interestingly, it was also observed that some molecules that internalize through clathrin-independent pathways (CD59, Bodipy-GM1, caveolin) also sequestered to the pericentrion upon sustained PKC activation, suggesting that PKC acted distal to the site of internalization of endocytic cargo. Together these results suggest that PKC regulates sequestration of recycling molecules into this compartment, the pericentrion.

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“…The position of the MM-associated proteins within the gradient was determined by immunoblotting for flotillin-1, a known MM marker 42,43 and showed flotillin-1 to be enriched in fractions 3-6, representing the 5-30% interface of the sucrose gradient ( Figure 1). In keeping with previous reports that the transferrin receptor is present in non-detergent-resistant membrane fractions and is solubilized under the above conditions, 44 this receptor was found in the soluble membrane fractions only (fraction 10; Figure 1).…”

Section: Flotillin-1 and Transferrin Receptor Distributionmentioning

confidence: 99%

Proteomic analysis of membrane microdomain-associated proteins in the dorsolateral prefrontal cortex in schizophrenia and bipolar disorder reveals alterations in LAMP, STXBP1 and BASP1 protein expression

et al. 2008

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The dorsolateral prefrontal cortex (dlpfc) is strongly implicated in the pathogenesis of schizophrenia (SCZ) and bipolar disorder (BPD) and, within this region, abnormalities in glutamatergic neurotransmission and synaptic function have been described. Proteins associated with these functions are enriched in membrane microdomains (MM). In the current study, we used two complementary proteomic methods, two-dimensional difference gel electrophoresis and one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis followed by reverse phase-liquid chromatography-tandem mass spectrometry (RP-LC-MS/MS) (gel separation liquid chromatography-tandem mass spectrometry (GeLC-MS/MS)) to assess protein expression in MM in pooled samples of dlpfc from SCZ, BPD and control cases (n = 10 per group) from the Stanley Foundation Brain series. We identified 16 proteins altered in one/both disorders using proteomic methods. We selected three proteins with roles in synaptic function (syntaxin-binding protein 1 (STXBP1), brain abundant membrane-attached signal protein 1 (BASP1) and limbic system-associated membrane protein (LAMP)) for validation by western blotting. This revealed significantly increased expression of these proteins in SCZ (STXBP1 (24% difference; P < 0.001), BASP1 (40% difference; P < 0.05) and LAMP (22% difference; P < 0.01)) and BPD (STXBP1 (31% difference; P < 0.001), BASP1 (23% difference; P < 0.01) and LAMP (20% difference; P < 0.01)) in the Stanley brain series (n = 20 per group). Further validation in dlpfc from the Harvard brain subseries (n = 10 per group) confirmed increased protein expression in SCZ of STXBP1 (18% difference; P < 0.0001), BASP1 (14% difference; P < 0.0001) but not LAMP (20% difference; P = 0.14). No significant differences in STXBP1, BASP1 or LAMP protein expression in BPD dlpfc were observed. This study, through proteomic assessments of MM in dlpfc and validation in two brain series, strongly implicates LAMP, STXBP1 and BASP1 in SCZ and supports the view of a neuritic and synaptic dysfunction in the neuropathology of SCZ.

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Ultrastructural localization of flotillin-1 to cholesterol-rich membrane microdomains, rafts, in rat brain tissue

Cited by 58 publications

References 26 publications

Enlargeosome, an Exocytic Vesicle Resistant to Nonionic Detergents, Undergoes Endocytosis via a Nonacidic Route

Enlargeosome, an Exocytic Vesicle Resistant to Nonionic Detergents, Undergoes Endocytosis via a Nonacidic Route

Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C

Proteomic analysis of membrane microdomain-associated proteins in the dorsolateral prefrontal cortex in schizophrenia and bipolar disorder reveals alterations in LAMP, STXBP1 and BASP1 protein expression

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