Effects of lipid phase transition and membrane surface charge on the interfacial activation of phospholipase A2.
Phospholipase A2 (PLA2) enzymes act at the membrane-water interface to access their phospholipid substrate from the membrane. They are regulated by diverse factors, including the membrane charge, fluidity, mode of membrane binding (insertion, orientation), and allosteric conformational effects. Relative contributions of these factors to the complex kinetics of PLA2 activation are not well understood. Here we examine the effects of thermal phase transitions and the surface charge of phospholipid membranes on the activation of human pancreatic PLA2. The temperature dependence of the initial catalytic rate of PLA2 peaks around the lipid phase transition temperature (Tm) when Tm is not too far from physiological temperatures (30-40 degrees C), and the peak is higher in the presence of anionic membranes. High PLA2 activity can be induced by thermal perturbations of the membrane. Temperature-dependent fluorescence quenching experiments show that despite dramatic effects of the lipid phase transition on PLA2 activity, the membrane insertion depth of PLA2 increases only modestly above Tm. The data show that membrane structural disorder, and not the depth of membrane insertion, plays a major role in PLA2 activity.
Ray
Supriyo
S
Biomolecular Science Center, Burnett College of Biomedical Sciences, University of Central Florida, 12722 Research Parkway, Orlando, Florida 32826, USA.
Scott
Jennifer L
JL
Tatulian
Suren A
SA
D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D008563:Membrane Lipids; D044367:Phase Transition; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D054467:Phospholipases A2 / Q000378:metabolism*; D013696:Temperature; D053835:Unilamellar Liposomes
D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D053835:Unilamellar Liposomes; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; C002773:dimyristoylphosphatidylglycerol; D054467:Phospholipases A2; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine