Hexadecylphosphocholine and 2-modified 1,3-diacylglycerols as effectors of phospholipase D.
The kinetic behaviour of phospholipase D (PLD) from cabbage has been studied in the presence of several substrate-like compounds such as hexadecylphosphocholine (HPC) and 1,3-didodecanoylglycero-2-phosphatides. 1,3-Didodecanoyl- glycero-2-phosphocholine (1,3-DiC12PC) was found being not cleft by PLD, whereas HPC is hydrolyzed by PLD with small rate. The plot of initial velocity vs. substrate concentration for HPC is more sigmoidal than those for the common substrate phosphatidylcholine (PC)/sodium dodecylsulfate (SDS) (1:0.5) or the short-chain 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DiC6PC). The anionic amphiphiles 1,3-didodecanoylglycero-2-sulfate and 1,3-didodecanoylglycero-2-phosphate act as activators of PLD towards PC similar to SDS. In contrast, 1,3-DiC12PC shows inhibitory properties with an increase in the sigmoidicity of the initial velocity as a function of substrate concentration in the PC/SDS assay. Also HPC inhibits the hydrolysis of PC/SDS, whereas it acts as activator or inhibitor in the hydrolysis of DiC6PC. The results suggest that PLD possesses two substrate-binding sites, where one binds substrate in function of an effector without catalytic activity while the other is the catalytic site. HPC and 1,3-DiC12PC are assumed to compete with the substrate for both binding sites with effects depending on the ratio of concentrations and affinities of substrates and effectors.
Dittrich
N
N
Department of Biochemistry/Biotechnology, Martin-Luther University Halle-Wittenberg, D-06099 Halle, Germany.
Haftendorn
R
R
Ulbrich-Hofmann
R
R
D001937:Brassica / Q000201:enzymology; D004075:Diglycerides / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D010713:Phosphatidylcholines / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D013379:Substrate Specificity; D013501:Surface-Active Agents
D004075:Diglycerides; D010713:Phosphatidylcholines; D013501:Surface-Active Agents; D010767:Phosphorylcholine; D012967:Sodium Dodecyl Sulfate; C039128:miltefosine; D010739:Phospholipase D