title: Kinetics and thermodynamics of annexin A1 binding to solid-supported membranes: a QCM study.
abstract: By means of the quartz crystal microbalance (QCM) technique, the interaction of annexin A1 with lipid membranes was quantified using solid-supported bilayers immobilized on gold electrodes deposited on 5 MHz quartz plates. Solid-supported lipid bilayers were composed of a first octanethiol monolayer chemisorbed on gold and a physisorbed phospholipid monolayer obtained from vesicle fusion. This experimental setup enabled us to determine for the first time rate constants and affinity constants of annexin A1 binding to phosphatidylserine-containing layers as a function of the calcium ion concentration in solution and the cholesterol content within the outer leaflet of the solid-supported bilayer. The results reveal that a decrease in Ca(2+) concentration from 1 mM to 100 microM significantly increases the rate of annexin A1 binding to the membrane independent of the cholesterol content. However, the presence of cholesterol in the membrane altered the affinity constants considerably. While the association constant decreases with decreasing Ca(2+) concentration in the case of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS) membranes lacking cholesterol, it remains high in the presence of cholesterol.
authors.affiliation: Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, 93040 Regensburg, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Kastl
authors.forename: Michaela
authors.initials: M
authors.lastname: Ross
authors.forename: Volker
authors.initials: V
authors.lastname: Gerke
authors.forename: Claudia
authors.initials: C
authors.lastname: Steinem
mesh_terms: D017302:Annexins / Q000378:metabolism*; D007700:Kinetics; D008567:Membranes, Artificial; D010743:Phospholipids / Q000378:metabolism; D011791:Quartz; D013816:Thermodynamics
chemical_list: D017302:Annexins; D008567:Membranes, Artificial; D010743:Phospholipids; D011791:Quartz