Yeast sphingosine-1-phosphate phosphatases: assay, expression, deletion, purification, and cellular localization by GFP tagging.
DHS-1-P phosphatases cloned from yeast represent novel lipid phosphatases, which were not thought to exist in yeast. Identification and characterization of YSR2 and YSR3 have demonstrated that the DHS-1-P phosphatase is an important mediator in the biosynthesis of sphingolipids and in the maintenance of the balance of signaling lipid molecules ceramide, sphingosine, and sphingosine-1-P. Methods introduced here for purification, activity assay, in vivo labeling, and cellular localization using GFP tagging are expected to facilitate our understanding of this enzyme.
Mao
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Department of Medicine, Medical University of South Carolina, Charleston 29425, USA.
Obeid
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D002451:Cell Compartmentation; D003001:Cloning, Molecular; D005800:Genes, Fungal; D018869:Heat-Shock Response; D016296:Mutagenesis; D009842:Oligopeptides; D019308:Palmitic Acid / Q000378:metabolism; D010455:Peptides; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D012694:Serine / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism; D014316:Tritium
D009842:Oligopeptides; D010455:Peptides; D010761:Phosphorus Radioisotopes; D029701:Saccharomyces cerevisiae Proteins; D013107:Sphingolipids; D014316:Tritium; C060504:dihydrosphingosine 1-phosphate; D019308:Palmitic Acid; D012694:Serine; C085706:FLAG peptide; C110252:LCB3 protein, S cerevisiae; C503018:YSR3 protein, S cerevisiae; D010744:Phosphoric Monoester Hydrolases; D013110:Sphingosine