title: Characterization of a sphingomyelinase activity in Saccharomyces cerevisiae.
abstract: Sphingomyelinases (SMase), which hydrolyze sphingolipids to yield ceramide, participate in signal transduction pathways in mammalian cells. Although yeast express many homologs of mammalian signaling proteins, SMase activity had not been previously demonstrated in yeast. In this study, we used an in vitro assay to characterize yeast SMase activity. Activity was detected in yeast membranes at both acid and neutral pH. The enzyme exhibited a requirement for magnesium or manganese, and was sensitive to detergents. The pI of the enzyme was approximately 5.9. SMase was separable from phospholipase D (PLD) activity, and was expressed at normal levels in yeast lacking expression of PLD1. While sphingosine and phytosphingosine inhibited growth, other sphingolipid metabolites had no effect on yeast growth. Intact yeast generate ceramide from exogenous sphingomyelin. These studies demonstrate that yeast express a membrane-localized neutral SMase activity.
authors.affiliation: Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Charleston 29425-2251, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Ella
authors.forename: C
authors.initials: C
authors.lastname: Qi
authors.forename: J W
authors.initials: JW
authors.lastname: Dolan
authors.forename: R P
authors.initials: RP
authors.lastname: Thompson
authors.forename: K E
authors.initials: KE
authors.lastname: Meier
mesh_terms: D000818:Animals; D002413:Cations, Divalent; D002460:Cell Line; D002518:Ceramides / Q000378:metabolism; D003902:Detergents / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008670:Metals; D010739:Phospholipase D / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D002518:Ceramides; D003902:Detergents; D008670:Metals; D013109:Sphingomyelins; D013108:Sphingomyelin Phosphodiesterase; D010739:Phospholipase D