Sphingosylphosphorylcholine as a novel calmodulin inhibitor.
S1P (sphingosine 1-phosphate) and SPC (sphingosylphosphorylcholine) have been recently recognized as important mediators of cell signalling, regulating basic cellular processes such as growth,differentiation, apoptosis, motility and Ca2+ homoeostasis.Interestingly, they can also act as first and second messengers. Although their activation of cell-surface G-protein-coupled receptors has been studied extensively, not much is known about heir intracellular mechanism of action, and their target proteins are yet to be identified. We hypothesized that these sphingolipids might bind to CaM (calmodulin), the ubiquitous intracellular Ca2+sensor. Binding assays utilizing intrinsic tyrosine fluorescence of the protein, dansyl-labelled CaM and surface plasmon resonance revealed that SPC binds to both apo- and Ca2+-saturated CaM selectively, when compared with the related lysophospholipid mediators S1P, LPA (lysophosphatidic acid) and LPC (lysophosphatidylcholine). Experiments carried out with the model CaM-binding domain melittin showed that SPC dissociates the CaM-target peptide complex, suggesting an inhibitory role. The functional effect of the interaction was examined on two target enzymes, phosphodiesterase and calcineurin, and SPC inhibited the Ca2+/CaM-dependent activity of both. Thus we propose that CaM might be an intracellular receptor for SPC, and raise the possibility of a novel endogenous regulation of CaM.
Kovacs
Erika
E
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Karolina ut 29, Budapest, H-1113, Hungary.
Liliom
Karoly
K
D000818:Animals; D001923:Brain Chemistry; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D002417:Cattle; D008081:Liposomes; D008823:Micelles; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000302:isolation & purification; D015398:Signal Transduction; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000502:physiology; D014443:Tyrosine / Q000032:analysis
D002147:Calmodulin; D008081:Liposomes; D008823:Micelles; C005356:sphingosine phosphorylcholine; D010767:Phosphorylcholine; D014443:Tyrosine; D013110:Sphingosine