Reversible adsorption and nonreversible insertion of Escherichia coli alpha-hemolysin into lipid bilayers.
Alpha-Hemolysin is an extracellular protein toxin (107 kDa) produced by some pathogenic strains of Escherichia coli. Although stable in aqueous medium, it can bind to lipid bilayers and produce membrane disruption in model and cell membranes. Previous studies had shown that toxin binding to the bilayer did not always lead to membrane lysis. In this paper, we find that alpha-hemolysin may bind the membranes in at least two ways, a reversible adsorption and an irreversible insertion. Reversibility is detected by the ability of liposome-bound toxin to induce hemolysis of added horse erythrocytes; insertion is accompanied by an increase in the protein intrinsic fluorescence. Toxin insertion does not necessarily lead to membrane lysis. Studies of alpha-hemolysin insertion into bilayers formed from a variety of single phospholipids, or binary mixtures of phospholipids, or of phospholipid and cholesterol, reveal that irreversible insertion is favored by fluid over gel states, by low over high cholesterol concentrations, by disordered liquid phases over gel or ordered liquid phases, and by gel over ordered liquid phases. These results are relevant to the mechanism of action of alpha-hemolysin and provide new insights into the membrane insertion of large proteins.
Bakás
L
L
Departamento de Bioquímica, Universidad del País Vasco, Bilbao, Spain.
Ostolaza
H
H
Vaz
W L
WL
Goñi
F M
FM
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000327:Adsorption; D001426:Bacterial Proteins / Q000737:chemistry*; D001427:Bacterial Toxins / Q000737:chemistry; D002784:Cholesterol / Q000737:chemistry; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers; D010713:Phosphatidylcholines / Q000737:chemistry*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D015060:1,2-Dipalmitoylphosphatidylcholine; D002784:Cholesterol; C010942:1,2-distearoyllecithin; C017251:1,2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine