Inhibition of caveolar uptake, SV40 infection, and beta1-integrin signaling by a nonnatural glycosphingolipid stereoisomer.
AuthorsSingh RD, Holicky EL, Cheng ZJ, Kim SY, Wheatley CL, Marks DL, Bittman R, Pagano RE
JournalJ Cell Biol
PubMed ID17371832
'Caveolar endocytosis is an important mechanism for the uptake of certain pathogens and toxins and also plays a role in the internalization of some plasma membrane (PM) lipids and proteins. However, the regulation of caveolar endocytosis is not well understood. We previously demonstrated that caveolar endocytosis and beta1-integrin signaling are ... More
Regulation of caveolar endocytosis by syntaxin 6-dependent delivery of membrane components to the cell surface.
AuthorsChoudhury A, Marks DL, Proctor KM, Gould GW, Pagano RE
JournalNat Cell Biol
PubMed ID16565709
'Caveolar endocytosis has an important function in the cellular uptake of some bacterial toxins, viruses and circulating proteins. However, the molecular machinery involved in regulating caveolar uptake is poorly defined. Here, we demonstrate that caveolar endocytosis is regulated by syntaxin 6, a target membrane soluble N-ethylmaleimide attachment protein receptor (t-SNARE) ... More
Selective caveolin-1-dependent endocytosis of glycosphingolipids.
AuthorsSingh RD, Puri V, Valiyaveettil JT, Marks DL, Bittman R, Pagano RE
JournalMol Biol Cell
PubMed ID12925761
'We studied the endocytosis of fluorescent glycosphingolipid (GSL) analogs in various cell types using pathway-specific inhibitors and colocalization studies with endocytic markers and DsRed caveolin-1 (cav-1). Based on inhibitor studies, all GSLs tested were internalized predominantly (>80%) by a clathrin-independent, caveolar-related mechanism, regardless of cell type. In addition, fluorescent lactosylceramide ... More
AuthorsSteinert S, Lee E, Tresset G, Zhang D, Hortsch R, Wetzel R, Hebbar S, Sundram JR, Kesavapany S, Boschke E, Kraut R,
JournalPLoS ONE
PubMed ID18716682
'BACKGROUND: The uptake and intracellular trafficking of sphingolipids, which self-associate into plasma membrane microdomains, is associated with many pathological conditions, including viral and toxin infection, lipid storage disease, and neurodegenerative disease. However, the means available to label the trafficking pathways of sphingolipids in live cells are extremely limited. In order ... More
Rab proteins mediate Golgi transport of caveola-internalized glycosphingolipids and correct lipid trafficking in Niemann-Pick C cells.
AuthorsChoudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL, Marks DL, Pagano RE
JournalJ Clin Invest
PubMed ID12070301
'We recently showed that human skin fibroblasts internalize fluorescent analogues of the glycosphingolipids lactosylceramide and globoside almost exclusively by a clathrin-independent mechanism involving caveolae. In contrast, a sphingomyelin analogue is internalized approximately equally via clathrin-dependent and caveolar routes. Here, we further characterized the caveolar pathway for glycosphingolipids, showing that Golgi ... More
Accumulation of glycosphingolipids in Niemann-Pick C disease disrupts endosomal transport.
Authorste Vruchte D, Lloyd-Evans E, Veldman RJ, Neville DC, Dwek RA, Platt FM, van Blitterswijk WJ, Sillence DJ
JournalJ Biol Chem
PubMed ID15078881
'Glycosphingolipids are endocytosed and targeted to the Golgi apparatus but are mistargeted to lysosomes in sphingolipid storage disorders. Substrate reduction therapy utilizes imino sugars to inhibit glucosylceramide synthase and potentially abrogate the effects of storage. Niemann-Pick type C (NPC) disease is a disorder of intracellular transport where glycosphingolipids (GSLs) and ... More
Cholesterol modulates membrane traffic along the endocytic pathway in sphingolipid-storage diseases.
AuthorsPuri V, Watanabe R, Dominguez M, Sun X, Wheatley CL, Marks DL, Pagano RE
JournalNat Cell Biol
PubMed ID10559968
Outlook for screening for sphingolipidoses.
AuthorsWinchester B
JournalLancet
PubMed ID10489940
Applications of BODIPY-sphingolipid analogs to study lipid traffic and metabolism in cells.
AuthorsPagano RE, Watanabe R, Wheatley C, Dominguez M
JournalMethods Enzymol
PubMed ID11070900
Use of Bodipy-labeled sphingolipid and cholesterol analogs to examine membrane microdomains in cells.
AuthorsMarks DL, Bittman R, Pagano RE,
JournalHistochem Cell Biol
PubMed ID18820942
Much evidence has accumulated to show that cellular membranes such as the plasma membrane, contain multiple
Glycosphingolipids in endocytic membrane transport.
AuthorsSillence DJ, Platt FM
JournalSemin Cell Dev Biol
PubMed ID15207831
Endocytosis leads to the internalisation of both lipids and proteins and their delivery to specific subcellular locations. This involves sorting processes that are not completely understood, but may involve interactions between lipids and proteins as well as pH and calcium gradients. This article discusses the importance of endocytosis in glycosphingolipid ... More
Use of fluorescent sphingolipid analogs to study lipid transport along the endocytic pathway.
AuthorsMarks DL, Singh RD, Choudhury A, Wheatley CL, Pagano RE
JournalMethods
PubMed ID15905102
Sphingolipids (SLs) are concentrated at the plasma membrane where they play important roles in cell-cell communication, host-pathogen interactions, and cell signaling events. Our laboratory has used fluorescent SL analogs and SL-binding toxins to elucidate mechanisms by which SLs are internalized by endocytosis and subsequently sorted and transported to various intracellular ... More
Cutting edge: negative regulation of immune synapse formation by anchoring lipid raft to cytoskeleton through Cbp-EBP50-ERM assembly.
AuthorsItoh K, Sakakibara M, Yamasaki S, Takeuchi A, Arase H, Miyazaki M, Nakajima N, Okada M, Saito T
JournalJ Immunol
PubMed ID11777944
Ag recognition by T lymphocytes induces immune synapse formation and recruitment of signaling molecules into a lipid raft. Cbp/PAG is a Csk-associated membrane adapter protein exclusively localized in a lipid raft. We identified NHERF/EBP50 as a Cbp-binding molecule, which connects the membrane raft and cytoskeleton by binding to both Cbp ... More
Cellular uptake of unconjugated TAT peptide involves clathrin-dependent endocytosis and heparan sulfate receptors.
Delivery of macromolecules mediated by protein transduction domains (PTDs) attracts a lot of interest due to its therapeutic and biotechnological potential. A major reevaluation of the mechanism of PTD-mediated internalization and the role of endocytosis in this mechanism has been recently initiated. Here, we demonstrate that the entry of TAT ... More
Cholesterol controls lipid endocytosis through Rab11.
AuthorsTakahashi M, Murate M, Fukuda M, Sato SB, Ohta A, Kobayashi T,
JournalMol Biol Cell
PubMed ID17475773
Cellular cholesterol increases when cells reach confluency in Chinese hamster ovary (CHO) cells. We examined the endocytosis of several lipid probes in subconfluent and confluent CHO cells. In subconfluent cells, fluorescent lipid probes including poly(ethylene glycol)derivatized cholesterol, 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3beta-ol, and fluorescent sphingomyelin analogs were internalized to pericentriolar recycling endosomes. This accumulation ... More
Glycolipid trafficking in Drosophila undergoes pathway switching in response to aberrant cholesterol levels.
AuthorsHortsch R, Lee E, Erathodiyil N, Hebbar S, Steinert S, Lee JY, Chua DS, Kraut R,
JournalMol Biol Cell
PubMed ID20053687
In lipid storage diseases, the intracellular trafficking of sphingolipids is altered by conditions of aberrant cholesterol accumulation. Drosophila has been used recently to model lipid storage diseases, but the effects of sterol accumulation on sphingolipid trafficking are not known in the fly, and the trafficking of sphingolipids in general has ... More
Deformation-induced lipid trafficking in alveolar epithelial cells.
AuthorsVlahakis NE, Schroeder MA, Pagano RE, Hubmayr RD
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID11290518
Mechanical ventilation with a high tidal volume results in lung injury that is characterized by blebbing and breaks both between and through alveolar epithelial cells. We developed an in vitro model to simulate ventilator-induced deformation of the alveolar basement membrane and to investigate, in a direct manner, epithelial cell responses ... More
The use of inhibitors to study endocytic pathways of gene carriers: optimization and pitfalls.
AuthorsVercauteren D, Vandenbroucke RE, Jones AT, Rejman J, Demeester J, De Smedt SC, Sanders NN, Braeckmans K,
JournalMol Ther
PubMed ID20010917
Nonviral gene complexes can enter mammalian cells through different endocytic pathways. For efficient optimization of the gene carrier it is important to profile its cellular uptake, because this largely determines its intracellular processing and subsequent transfection efficiency. Most of the current information on uptake of these gene-delivery vehicles is based ... More
Membrane traffic in sphingolipid storage diseases.
AuthorsPagano RE, Puri V, Dominguez M, Marks DL
JournalTraffic
PubMed ID11208071
In this review, we summarize our studies of membrane lipid transport in sphingolipid storage disease (SLSD) fibroblasts. We recently showed that several fluorescent SL analogs were internalized from the plasma membrane predominantly to the Golgi complex of normal cells, while in ten different SLSD cell types, these lipids accumulated in ... More
The lipoprotein receptor-related protein-1 (LRP) adapter protein GULP mediates trafficking of the LRP ligand prosaposin, leading to sphingolipid and free cholesterol accumulation in late endosomes and impaired efflux.
AuthorsKiss RS, Ma Z, Nakada-Tsukui K, Brugnera E, Vassiliou G, McBride HM, Ravichandran KS, Marcel YL
JournalJ Biol Chem
PubMed ID16497666
One of the conserved functional pathways linked to engulfment of apoptotic corpses involves two membrane proteins low density lipoprotein receptor-related protein-1 (LRP) and ABCA1 and the LRP adapter protein GULP. Because LRP and ABCA1 play roles in cellular lipid trafficking and efflux, here we addressed whether the third member, the ... More
Clathrin-dependent and -independent internalization of plasma membrane sphingolipids initiates two Golgi targeting pathways.
AuthorsPuri V, Watanabe R, Singh RD, Dominguez M, Brown JC, Wheatley CL, Marks DL, Pagano RE
JournalJ Cell Biol
PubMed ID11481344
Sphingolipids (SLs) are plasma membrane constituents in eukaryotic cells which play important roles in a wide variety of cellular functions. However, little is known about the mechanisms of their internalization from the plasma membrane or subsequent intracellular targeting. We have begun to study these issues in human skin fibroblasts using ... More
Glucosylceramide modulates membrane traffic along the endocytic pathway.
AuthorsSillence DJ, Puri V, Marks DL, Butters TD, Dwek RA, Pagano RE, Platt FM
JournalJ Lipid Res
PubMed ID12401882
Glycosphingolipids are endocytosed and targeted to the Golgi apparatus, but are mistargeted to lysosomes in numerous sphingolipidoses. Substrate reduction therapy utilizes imino sugars to inhibit glucosylceramide synthase and potentially abrogate the effects of storage. Gaucher disease is a hereditary deficiency in glucocerebrosidase leading to glucosylceramide accumulation; however, Gaucher fibroblasts exhibited ... More
Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis.
AuthorsRejman J, Oberle V, Zuhorn IS, Hoekstra D
JournalBiochem J
PubMed ID14505488
Non-phagocytic eukaryotic cells can internalize particles <1 microm in size, encompassing pathogens, liposomes for drug delivery or lipoplexes applied in gene delivery. In the present study, we have investigated the effect of particle size on the pathway of entry and subsequent intracellular fate in non-phagocytic B16 cells, using a range ... More
Internalization and sorting of plasma membrane sphingolipid analogues in differentiating oligodendrocytes.
AuthorsWatanabe R, Asakura K, Rodriguez M, Pagano RE
JournalJ Neurochem
PubMed ID10501180
We studied the formation of early endosomes in differentiating oligodendrocytes and type-2 astrocytes, which are derived from common precursor cells in rat neonates, using fluorescent analogues of lactosylceramide (LacCer) and sulfatide labeled with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene++ +-3-pentanoic acid (BODIPY FL C5). These sphingolipid analogues exhibit a concentration-dependent shift in their fluorescence emission ... More
Internalization and sorting of a fluorescent analogue of glucosylceramide to the Golgi apparatus of human skin fibroblasts: utilization of endocytic and nonendocytic transport mechanisms.
AuthorsMartin OC, Pagano RE
JournalJ Cell Biol
PubMed ID8188745
We examined the uptake and intracellular transport of the fluorescent glucosylceramide analogue N-[5-(5,7-dimethyl BODIPYTM)-1-pentanoyl]-glucosyl sphingosine (C5-DMB-GlcCer) in human skin fibroblasts, and we compared its behavior to that of the corresponding fluorescent analogues of sphingomyelin, galactosylceramide, and lactosylceramide. All four fluorescent analogues were readily transferred from defatted BSA to the plasma ... More
Abnormal transport along the lysosomal pathway in mucolipidosis, type IV disease.
AuthorsChen CS, Bach G, Pagano RE
JournalProc Natl Acad Sci U S A
PubMed ID9600972
Mucolipidosis, type IV (ML-IV) is an autosomal recessive storage disease that is characterized by lysosomal accumulation of sphingolipids, phospholipids, and acid mucopolysaccharides. Unlike most other storage diseases, the lysosomal hydrolases participating in the catabolism of the stored molecules appear to be normal. In the present study, we examined the hypothesis ... More
Broad screening test for sphingolipid-storage diseases.
AuthorsChen CS, Patterson MC, Wheatley CL, O'Brien JF, Pagano RE
JournalLancet
PubMed ID10489949
BACKGROUND: Lipid-storage diseases are collectively important because they cause substantial morbidity and mortality, and because they may present as dementia, major psychiatric illness, developmental delay, or cerebral palsy. At present, no single assay can be used as an initial general screen for lipid-storage diseases. METHODS: We used a fluorescent analogue ... More
Glycosphingolipids internalized via caveolar-related endocytosis rapidly merge with the clathrin pathway in early endosomes and form microdomains for recycling.
AuthorsSharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE
JournalJ Biol Chem
PubMed ID12482757
We have previously demonstrated that glycosphingolipids are internalized from the plasma membrane of human skin fibroblasts by a clathrin-independent, caveolar-related mechanism and are subsequently transported to the Golgi apparatus by a process that is dependent on microtubules, phosphatidylinositol 3-kinase, Rab7, and Rab9. Here we characterized the early steps of intracellular ... More
Niemann-Pick C variant detection by altered sphingolipid trafficking and correlation with mutations within a specific domain of NPC1.
AuthorsSun X, Marks DL, Park WD, Wheatley CL, Puri V, O'Brien JF, Kraft DL, Lundquist PA, Patterson MC, Pagano RE, Snow K
JournalAm J Hum Genet
PubMed ID11349231
Niemann-Pick disease type C (NPC) is a fatal, autosomal recessive lipidosis characterized by lysosomal accumulation of unesterified cholesterol and multiple neurological symptoms, such as vertical supranuclear ophthalmoplegia, progressive ataxia, and dementia. More than 90% of cases of NPC are due to a defect in Niemann-Pick C1 (NPC1), a late endosomal, ... More
TRIM72 modulates caveolar endocytosis in repair of lung cells.
Authors
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID26637632
The SPPL3-Defined Glycosphingolipid Repertoire Orchestrates HLA Class I-Mediated Immune Responses.
Authors
JournalImmunity
PubMed ID33271119
Age-related increase in caveolin-1 expression facilitates cell-to-cell transmission of α-synuclein in neurons.