Measurement of mammalian sphingosine-1-phosphate phosphohydrolase activity in vitro and in vivo.
Sphingolipid metabolites have emerged as key players in diverse processes including cell migration, growth, and apoptosis. Ceramide and sphingosine typically inhibit cell growth and induce apoptosis, while sphingosine-1-phosphate (S1P) promotes cell growth, inhibits apoptosis, and induces cell migration. Thus, enzymes that regulate the levels of these sphingolipid metabolites are of critical importance to understanding cell fate. There are two known mammalian isoforms of S1P phosphohydrolases (SPP1 and SPP2) that reversibly degrade S1P to sphingosine. This chapter discusses the importance of SPPs and describes assays that can be used to measure the activity of these two specific S1P phosphohydrolases in cells and cell lysates.
Maceyka
Michael
M
Department of Biochemistry and Molecular Biology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.
Milstien
Sheldon
S
Spiegel
Sarah
S
D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008246:Lysophospholipids / Q000378:metabolism; D008322:Mammals; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D008246:Lysophospholipids; D008565:Membrane Proteins; C060506:sphingosine 1-phosphate; C401298:sphingosine-1-phosphate phosphatase; D010744:Phosphoric Monoester Hydrolases; D013110:Sphingosine; D005033:Ethylmaleimide