Reconstitution of ATP- and Cytosol-dependent Transport of de Novo Synthesized Ceramide to the Site of Sphingomyelin Synthesis in Semi-intact Cells

Funakoshi, Tomoko; Yasuda, Satoshi; Fukasawa, Masayoshi; Nishijima, Masahiro; Hanada, Kentaro

doi:10.1074/jbc.m004470200

Cited by 56 publications

(80 citation statements)

References 35 publications

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“…An in vitro reconstitu- tion assay system within semi-intact CHO cells was shown to reproduce various aspects of the ER-to-Golgi trafficking of ceramide (9,13,17,18). This reconstitution method allowed us to adjust the quantity of CERT proteins added to the assay system.…”

Section: Phosphorylation At Ser-315 Up-regulated Cert-mediated Transpmentioning

confidence: 99%

“…Assay of the ER-to-Golgi Transport of Ceramide in Semi-intact Cells-The assay was performed as described previously (9,17,18 Cell-free Assay of the Intermembrane Transfer of Ceramide-A cell-free assay of the intermembrane transfer of ceramide was performed with modifications according to a previously reported method (9,11). Briefly, donor and acceptor liposomes composed of POPC/POPE/dioleoyl-sn-glycerol-3-phosphoethanolamine-N-lactosyl (ammonium salt)/[ 14 C]ceramide (55 mCi/mmol) (64:16:40:1, mol/mol) and POPC/POPE (4:1, mol/ mol), respectively, were formed in buffer C (20 mM HepesNaOH (pH 7.4) containing 50 mM NaCl and 1 mM EDTA) by probe-type sonication.…”

Section: Construction Of a Plasmid For Bacterial Expression Of A Tranmentioning

confidence: 99%

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Phosphoregulation of the Ceramide Transport Protein CERT at Serine 315 in the Interaction with VAMP-associated Protein (VAP) for Inter-organelle Trafficking of Ceramide in Mammalian Cells

Kumagai

Kawano‐Kawada²,

Hanada

2014

Journal of Biological Chemistry

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Background: CERT transports ceramide from the ER to the Golgi apparatus and is supported by its FFAT motif-dependent binding to the ER membrane protein VAP. Results: CERT is phosphorylated at Ser-315 near the FFAT motif. Conclusion:The phosphorylation strengthens the CERT-VAP-A interaction to up-regulate ceramide trafficking. Significance: This provides new insights into molecular mechanisms underlying functional regulation of FFAT-containing proteins for the inter-organelle lipid trafficking.

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Section: Phosphorylation At Ser-315 Up-regulated Cert-mediated Transpmentioning

confidence: 99%

Section: Construction Of a Plasmid For Bacterial Expression Of A Tranmentioning

confidence: 99%

Phosphoregulation of the Ceramide Transport Protein CERT at Serine 315 in the Interaction with VAMP-associated Protein (VAP) for Inter-organelle Trafficking of Ceramide in Mammalian Cells

Kumagai

Kawano‐Kawada²,

Hanada

2014

Journal of Biological Chemistry

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“…33 The nonvescicular transfer of ceramide from the ER to the Golgi requires ATP. 34,35 The ATP-dependent step for ceramide trafficking might be attributable to the synthesis of PI4P in the Golgi. Alternatively, ATP may allow the recycling of CERT after its binding to PI4P or VAP or it could energize an ATP-dependent translocase that moves ceramide from the cytosolic side of the Golgi to the luminal side where the catalytic domain of SMS is located.…”

Section: Discovery Of a Ceramide Transfer Protein With A Key Role In mentioning

confidence: 99%

Tales and Mysteries of the Enigmatic Sphingomyelin Synthase Family

Holthuis

Luberto

2010

Advances in Experimental Medicine and Biology

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In the last five years tremendous progress has been made toward the understanding of the mechanisms that govern sphingomyelin (SM) synthesis in animal cells. In line with the complexity of most biological processes, also in the case of SM biosynthesis, the more we learn the more enigmatic and finely tuned the system appears. Therefore with this review we aim first, at highlighting the most significant discoveries that advanced our knowledge and understanding of SM biosynthesis, starting from the discovery of SM to the identification of the enzymes responsible for its production; and second, at discussing old and new riddles that such discoveries pose to current investigators.

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“…This necessitates the transport of ceramide from the ER to the Golgi complex. The transport of ceramide is achieved mostly through an active ATP-dependent non-vesicular mode and by a minor ATP-independent pathway that may include vesicular transport [36,[112][113][114]. Hanada and his colleagues recently discovered a novel protein that is responsible for the former transport mode.…”

Section: Microdomains and Membrane Integritymentioning

confidence: 99%

Sphingolipids and membrane biology as determined from genetic models

Rao

Acharya

2008

Prostaglandins & Other Lipid Mediators

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The importance of sphingolipids in membrane biology was appreciated early in the twentieth century when several human inborn errors of metabolism were linked to defects in sphingolipid degradation. The past two decades have seen an explosion of information linking sphingolipids with cellular processes. Studies have unraveled mechanistic details of the sphingolipid metabolic pathways, and these findings are being exploited in the development of novel therapies, some now in clinical trials. Pioneering work in yeast has laid the foundation for identifying genes encoding the enzymes of the pathways. The advent of the era of genomics and bioinformatics has led to the identification of homologous genes in other species and the subsequent creation of animal knock-out lines for these genes. Discoveries from these efforts have re-kindled interest in the role of sphingolipids in membrane biology. This review highlights some of the recent advances in understanding sphingolipids' roles in membrane biology as determined from genetic models.

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Reconstitution of ATP- and Cytosol-dependent Transport of de Novo Synthesized Ceramide to the Site of Sphingomyelin Synthesis in Semi-intact Cells

Cited by 56 publications

References 35 publications

Phosphoregulation of the Ceramide Transport Protein CERT at Serine 315 in the Interaction with VAMP-associated Protein (VAP) for Inter-organelle Trafficking of Ceramide in Mammalian Cells

Phosphoregulation of the Ceramide Transport Protein CERT at Serine 315 in the Interaction with VAMP-associated Protein (VAP) for Inter-organelle Trafficking of Ceramide in Mammalian Cells

Tales and Mysteries of the Enigmatic Sphingomyelin Synthase Family

Sphingolipids and membrane biology as determined from genetic models

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