The monolayer properties of Atriopeptin III (AP III), a potent natural hypotensive agent, have been studied at the air/water interface. The surface pressure-molecular area isotherms of the AP III monolayer and mixed layers of the peptide with neutral and anionic phospholipids have been measured. Circular dichroism spectra of the transferred layers (Langmuir-Blodgett films) of the peptide, and those of the mixed layers of the peptide and lipids, on quartz substrates have been studied. The spectra show that the peptide adopts largely an alpha-helix structure in the presence of negatively charged lipid. This has been confirmed by attenuated total reflection Fourier transform-infrared spectroscopy.