Modulation of phospholipases A2 and C activities against dilauroylphosphorylcholine in mixed monolayers with semisynthetic derivatives of ganglioside and sphingosine.
Most of the semisynthetic ganglioside and sphingosine derivatives studied here decreased the rate as well as the extent of hydrolysis of monomolecular layers of dilauroylphosphorylcholine (dlPC) by both phospholipase A2 (PLA2) and C (PLC). For PLA2, the rate of enzymatic activity was inversely correlated (p < 0.001) with the duration of the latency period of the enzymatic reaction. The correlation between the rate of activity and the latency period was not statistically significant for PLC. The potency to inhibit PLA2 and PLC was not significantly correlated with the presence of specific chemical groups. Also, the inhibitory effect is not correlated to changes of the substrate intermolecular spacing or surface potential caused by the sphingolipids (SLs). Conversely, for PLA2 the variation of the kinetic parameters of the reaction with the molecular polarization vector of the SLs are statistically significant (p < 0.01). The rate of phospholipid degradation by PLA2 decreased, and the lag times tended to become longer, with increasing values of the SLs' polarization vector. The kinetic parameters of the reaction with PLC did not show statistically significant correlation with the polarization vector of the SLs. Our results suggest that the configuration of the electrostatic field across the interface is more important than are formal charges or specific chemical moieties in modulating the activity of PLA2. Inhibition of phospholipase activities of these types by specific SLs or their metabolites may be important as supramolecular regulatory steps at the membrane level of the amplification of lipid-mediated signaling pathways.
Perillo
M A
MA
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0614.
Guidotti
A
A
Costa
E
E
Yu
R K
RK
Maggio
B
B
D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000378:metabolism; D004563:Electrochemistry; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008954:Models, Biological; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D015398:Signal Transduction; D013110:Sphingosine / Q000737:chemistry / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism*
D005732:Gangliosides; D010713:Phosphatidylcholines; C015153:1,2-dilauroylphosphatidylcholine; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases; D013110:Sphingosine