Reconstitution assay system for ceramide transport with semi-intact cells.
The intracellular transport of lipids from the sites of their synthesis to their appropriate destination is a critical step for lipid metabolism. One well-defined inter-organelle lipid movement is the transport of ceramide by ceramide transport protein (CERT). Ceramide, a key intermediate for both sphingomyelin and glycosphingolipids, is synthesized at the endoplasmic reticulum and delivered to the Golgi apparatus to be converted to sphingomyelin. CERT delivers ceramide from the ER to the Golgi apparatus in a non-vesicular and ATP-dependent manner. This chapter describes a reconstitution assay system for ceramide transport with semi-intact cells, which is useful for the study of the CERT-mediated inter-organelle transport of ceramide.
Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan.
Keigo
K
Kumagai
Masahiro
M
Nishijima
Kentaro
K
Hanada
D000818:Animals; D001693:Biological Transport, Active; D002021:Buffers; D016466:CHO Cells; D002352:Carrier Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D018929:Cell Culture Techniques; D002457:Cell Extracts / Q000302:isolation & purification; D002518:Ceramides / Q000378:metabolism*; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism
D002021:Buffers; D002352:Carrier Proteins; D002457:Cell Extracts; D002518:Ceramides; D011993:Recombinant Fusion Proteins; C119090:CERT1 protein, human; D017346:Protein Serine-Threonine Kinases