Electrostatic and hydrophobic interactions are involved in factor Va binding to membranes containing acidic...

The interaction of factor Va with phospholipid monolayers was studied using alternating current polarography. For these studies a hanging mercury drop electrode is positioned in contact with the monolayer at the air-monolayer interface. Factor Va introduced into the aqueous subphase beneath the monolayer caused alterations in the capacitance of the circuit. When factor Va was introduced beneath monolayers containing 13-25% mole fraction phosphatidylserine (PS), 87-75% mole fraction phosphatidylcholine (PC), a significant capacitance increase occurred, which is interpreted to be the result of the penetration of the factor Va molecules through the monolayer. No penetration or associated capacitance changes were observed with monolayers composed of pure PC or pure PS. Polarography experiments were also conducted with the electrode placed in the aqueous subphase to measure the binding of factor Va to pure PS monolayers. This approach has allowed detection of factor Va (at nM concentrations) adsorption to a pure PS monolayer. Thus the membrane adsorption process can be distinguished from factor Va penetration through the monolayer. These results suggest that there are two types of interaction of the factor Va molecule with PS-containing monolayers. The interaction with the PS component of the monolayer is essential for binding the factor Va (at nM concentrations) that occurs by surface adsorption. For mixed PC/PS monolayers, PS adsorption is followed by penetration of the protein through the membrane.