The lytic activity of the bee venom peptide melittin is strongly reduced by the presence of negatively charged phospholipids or chloroplast galactolipids in the membranes of phosphatidylcholine large unilamellar vesicles.
We have investigated the dependence of the lytic activity of the bee venom peptide melittin on the lipid composition of its target membrane. The lysis of large unilamellar liposomes, measured as loss of the fluorescent dye carboxyfluorescein, in the presence of melittin was strongly reduced when the negatively charged lipids phosphatidylglycerol (PG) or phosphatidylserine (PS), or the plant chloroplast lipids monogalactosyldiacylglycerol (MGDG) or digalactosyldiacylglycerol (DGDG) were incorporated into egg phosphatidylcholine (EPC) membranes. This reduction was evident at concentrations below 10 wt% of the additional lipids. It was not due to reduced binding of melittin to the vesicles. It was also not related to a reduced insertion depth of the peptide into the bilayer, as shown by quenching of the intrinsic tryptophan fluorescence of the peptide by the aqueous quencher sodium nitrate. Fourier transform infrared spectroscopy (FTIR) revealed specific interactions of the peptide with the headgroups of the inhibitory lipids. The phosphate peak in PG was shifted by two wavenumbers after the addition of melittin. There was no shift in EPC or PS. Instead, in PS the COO- peak was strongly distorted in the presence of melittin. These data indicate ionic interactions between the basic peptide and the negative charges on the membrane surface. The galactolipids are uncharged. Here the evidence points to hydrogen bonding between melittin and OH-groups of the sugar headgroups. Liposomes containing DGDG were the only case where we found evidence for changes in fatty acyl chain motion due to the presence of melittin, from the CH2-scissoring peaks.
Hincha
D K
DK
Institut für Pflanzenphysiologie und Mikrobiologie, Freie Universität, Berlin, Germany.
Crowe
J H
JH
D001514:Bee Venoms; D002736:Chloroplasts / Q000502:physiology; D038983:Galactolipids; D006017:Glycolipids; D007700:Kinetics; D008081:Liposomes; D008555:Melitten / Q000037:antagonists & inhibitors; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D017550:Spectroscopy, Fourier Transform Infrared; D018724:Spinacia oleracea
D001514:Bee Venoms; D038983:Galactolipids; D006017:Glycolipids; D008081:Liposomes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D008555:Melitten