Proteins at air–water interfaces studied using external reflection circular dichroism

In this report we describe the first attempts to record external reflection circular dichroism (ERCD) spectra of beta-lactoglobulin solutions. It is shown that the accumulated proteins at and near the air-water interface can be detected using ERCD and that the signals obtained contain information on the conformational properties and concentration of the proteins residing at the interface. The local protein concentration and its conformation are in full agreement with previous observations using external reflection infrared spectroscopy. The ERCD signals are dominated by linear dichroism (LD) due to non-ideal behavior of the instrumental optics, but can be explained for using the theoretical description of chiral reflection. This allows the analysis of ERCD spectra of protein solutions. The measured ERCD signals are described accurately in the region between 190 and 220 nm, but poor resemblance is obtained at higher wavelengths. We are however confident that improvement of experimental conditions and theoretical description will allow that in the near future, external reflection circular dichroism (CD) can be a valuable tool that complements the application of external reflection infrared spectroscopy to study interfacial systems.