FIG. 2.
Cross-linked LFN cannot translocate across artificial membranes. (A) Schematic representation of translocation as assessed in planar lipid bilayers. Binding and translocation of LFN were monitored by conductance. Pores initially form in membranes after addition of heptameric PA63. Addition of LFN rapidly lowers conductance, mimicking closed channels. Translocation, which is initiated by increasing the voltage to +50 mV, promotes passage of LFN to the trans side of the membrane. (Modified from reference 13 with permission of the publisher.) (B) PA pores were formed in membranes at a transmembrane voltage of +20 mV and incubated in the presence of 10 nM monomeric or multimeric LFN constructs. Ion conductance was measured during LFN incubation and perfusion of the cis chamber. (C) PA channels blocked with monomeric or multimeric LFN were subjected to an increase in transmembrane voltage to +50 mV, and ion conductance was monitored until a steady state was reached. WT, wild type.