Complex formation of empedopeptin with peptidoglycan precursors lipid I, lipid II, and C55-PP and estimation of binding stoichiometry. Lipid I (A), lipid II (B), and C55-PP (C) were incubated with empedopeptin (EMP) at the indicated molar ratios in the presence of 1.25 mm calcium. The stable complex of empedopeptin with lipid I or lipid II remains close to the application spot, whereas free lipids migrate to the indicated positions. C55-PP was not trapped in a complex up to a 4-fold molar excess of the antibiotic. Rf(lipid I), 0.33; Rf(lipid I-EMP complex), 0.17; Rf(lipid II), 0.3; Rf(lipid II-EMP complex), 0.16; Rf(C55-PP), 0.37.