title: Formation of domains of cationic or anionic lipids in binary lipid mixtures increases the electrostatic coupling strength of water-soluble proteins to supported bilayers.
abstract: The electrostatic binding strength of water-soluble proteins having either an excess positive (cytochrome c) or negative (beta-lactoglobulin) electric charge to oppositely charged supported planar bilayers (SPBs) was studied as a function of the bilayer phase state (fluid or gel phase) by IR-ATR spectroscopy. The bilayer consisted of mixtures of zwitterionic DEPC with either cationic DMTAP or anionic DMPG. We observed drastic differences in the binding strength of both proteins for the two bilayer phase states, with the gel phase exhibiting a higher binding strength than the fluid phase, under conditions where the two lipid components had different hydrophobic chain lengths resulting in a nonideal mixing behavior. In addition, for beta-lactoglobulin we observed a strong binding to a gel phase SPB comprising DEPC/DMTAP, while raising the temperature of the SPB above the chain melting transition temperature of the mixture resulted in a complete unbinding of the protein. In contrast, for DMPC/DMTAP having the same cationic charge content but no hydrophobic chain mismatch, no phase-dependent coupling strength of the protein to the SPB was observed. Our results suggest that the formation of charge-enriched domains by partial demixing of the bilayer lipids at the transition to the gel state is crucial for modulation of the protein binding strength to the SPB, while the intrinsic charge of the solid support surface is of minor importance.
authors.affiliation: Universität Würzburg, Physikalisches Institut EP-5, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Käsbauer
authors.forename: T M
authors.initials: TM
authors.lastname: Bayerl
mesh_terms: D000818:Animals; D000838:Anions / Q000737:chemistry / Q000378:metabolism; D002412:Cations / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry* / Q000378:metabolism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D007782:Lactoglobulins / Q000737:chemistry* / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D009226:Myristates / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D011485:Protein Binding; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012995:Solubility; D013055:Spectrophotometry, Infrared; D055672:Static Electricity; D014867:Water / Q000737:chemistry
chemical_list: C117241:1,2-dimyristoyl-3-trimethylammonium propane; D000838:Anions; D002412:Cations; D003574:Cytochrome c Group; D007782:Lactoglobulins; D008051:Lipid Bilayers; D009226:Myristates; D010713:Phosphatidylcholines; D000644:Quaternary Ammonium Compounds; D014867:Water; C017251:1,2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine