title: Combination of C(17) sphingoid base homologues and mass spectrometry analysis as a new approach to study sphingolipid metabolism.
abstract: In recent years, sphingolipid metabolites ceramide, sphingosine, and sphingosine-1-phosphate have emerged as important second messengers in addition to their role as precursors of biomembrane components. The investigation of these sphingolipid metabolites requires the development of new, more sensitive methods for assaying the enzymes involved in their production. This chapter describes the utilization of mass spectrometry technology in combination with nonnaturally occurring C(17) sphingoid bases in the in vitro assays of two of the enzymes of the sphingolipid pathway, ceramide synthase and sphingosine kinase. These new in vitro methods provide high sensitivity and extreme accuracy even when crude extracts are used as enzyme sources.
authors.affiliation: Department of Medicine, Medical University of South Carolina, Charleston, South Carolina, USA.
authors.forename: Stefka
authors.initials: S
authors.lastname: Spassieva
authors.forename: Jacek
authors.initials: J
authors.lastname: Bielawski
authors.forename: Viviana
authors.initials: V
authors.lastname: Anelli
authors.forename: Lina M
authors.initials: LM
authors.lastname: Obeid
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005227:Fatty Acids / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000302:isolation & purification / Q000378:metabolism*; D013107:Sphingolipids / Q000302:isolation & purification* / Q000378:metabolism*; D000099157:Sphingosine Kinase
chemical_list: D005227:Fatty Acids; D010088:Oxidoreductases; D017853:Phosphotransferases (Alcohol Group Acceptor); D013107:Sphingolipids; D000099157:Sphingosine Kinase; C108138:dihydroceramide desaturase