Association of lysozyme with phospholipid vesicles is accompanied by membrane surface dehydration.
Lysozyme is a globular protein which is known to bind to negatively charged phospholipid vesicles. In order to study the relationship between binding of the protein and the subsequent destabilization of the phospholipid vesicles a set of experiments was performed using phospholipid monolayers and vesicles. Using microelectrophoresis the binding of lysozyme to phospholipid vesicles made of PS was determined. At low ionic strength and mild acidic pH of the solution lysozyme reduced the magnitude of the negative zeta potential of PS vesicles at lower concentrations compared to neutral pH and high ionic strength. In contrast, the bound fraction of lysozyme to PS vesicles was nearly constant at acidic and neutral pH. At low pH, the binding of lysozyme was accompanied by a strong aggregation of the vesicles. Lysozyme binding to PS vesicles is accompanied by its penetration into the PL monolayer. This was measured by surface tension and film balance measurements at low pH and low ionic strength. The interaction of lysozyme with negatively charged vesicles lead to a decrease of the vesicle surface hydration as measured by the shift of the emission peak of the fluorescent probe DPE. The binding of bis-ANS increased at low pH after addition of lysozyme to the vesicles. This indicates that more hydrophobic patches of the lysozyme-PS complex are exposed at low pH. At low pH the binding process of lysozyme to PS vesicles was followed by an extensive intermixing of phospholipids between the aggregated vesicles, accompanied by a massive leakage of the aqueous content of vesicles.
University of Leipzig, Institute for Medical Physics and Biophysics, FRG. zsco@medizin.uni-leipzig.de
O
O
Zschörnig
G
G
Paasche
C
C
Thieme
N
N
Korb
A
A
Fahrwald
K
K
Arnold
D000817:Anilino Naphthalenesulfonates / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D004983:Ethanolamines / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008027:Light; D009113:Muramidase / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence
D000817:Anilino Naphthalenesulfonates; D004983:Ethanolamines; D005456:Fluorescent Dyes; D007477:Ions; C120418:N-benzyl-1,2-diphenylethanolamine; D010743:Phospholipids; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D009113:Muramidase