One member of a prototypical class of antimicrobial oligomers was used to study pore formation in cardiolipin-rich membranes. Both vesicle dye-leakage assays and small-angle X-ray scattering were used to study bilayer remodeling. The results indicate that the presence of negative intrinsic curvature lipids is essential for pore formation by this class of molecules: In Gram-positive bacteria, cardiolipin and divalent metal cations like Ca2+ and Mg2+ are needed. This is consistent with the rote of phosphatidylethanolamine (PE) lipid in Gram-negative bacteria, where antimicrobial activity is dependent on the negative intrinsic curvature of PE rather than a specific interaction with PE.