Interaction of myelin basic protein with artificial membranes. Parameters governing binding, aggregation and dissociation.
The interaction of myelin basic protein (MBP) with large unilamellar vesicles, composed of phosphatidylserine (PtdSer), phosphatidylserine/phosphatidylcholine (PtdSer/Ole2GroPCho) and phosphatidylcholine/cholesterol (Ole2GroPCho/cholesterol) was examined. Binding of MBP to the bilayers as well as the kinetics of this process were determined by a resonance energy transfer procedure. The ability of the protein to aggregate the vesicles subsequently was monitored continuously by absorbance measurements. The interaction was further characterized by determining the ability of MBP to induce membrane perturbations, as reflected by release of aqueous vesicle contents, and lipid mixing. The results demonstrate that Ole2GroPCho inhibits, while PtdSer and cholesterol strongly facilitate MBP-induced membrane aggregation. Furthermore, binding of MBP to vesicles and the subsequent aggregation event are separate processes, i.e. the extent of binding does not necessarily reflect the aggregation susceptibility. Overall, aggregation appears to be the rate-limiting step. Interaction of MBP with PtdSer bilayers results in a limited degree of lipid mixing, which is accompanied by extensive release of vesicle contents. For all other compositions, no lipid mixing occurs, while cholesterol effectively prevents release of vesicle contents. pH-dependent experiments indicate distinct mechanisms to be operative in MBP-induced aggregation of PtdSer and Ole2GroPCho/cholesterol bilayers. At neutral pH, protein-protein interactions appear relevant, while at acidic pH intervesicular bridges, established by monomers that may cause aggregation of PtdSer vesicles, but not of Ole2GroPCho/cholesterol vesicles. The observation that divalent cations reverse MBP-induced vesicle aggregation may have physiological relevance.
ter Beest
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MB
Laboratory of Physiological Chemistry, University of Groningen, The Netherlands.
Hoekstra
D
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D002118:Calcium / Q000494:pharmacology; D002784:Cholesterol / Q000378:metabolism / Q000494:pharmacology; D004735:Energy Transfer; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism*; D046911:Macromolecular Substances; D004676:Myelin Basic Protein / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism / Q000494:pharmacology; D010718:Phosphatidylserines / Q000378:metabolism
D008051:Lipid Bilayers; D008081:Liposomes; D046911:Macromolecular Substances; D004676:Myelin Basic Protein; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine; D002118:Calcium