Polarized infrared spectra of melittin incorporated into macroscopically oriented lipid membranes are reported. From the linear dichroism of the amide I and amide II vibrational bands, the spatial orientation of the melittin helices was determined as being preferentially parallel to the membrane normal, under our experimental condition of low water content and an ordered lipid phase. Considering the various models for the orientation of melittin in lipid membranes proposed in the literature, we conclude that our data are in accord with an arrangement whereby the hydrophobic part of the polypeptide either spans the bilayer in the form of two bent helix segments, or is folded back within one monolayer in the form of a wedge.