Electrostatic sensor for identifying interactions between peptides and bacterial membranes.

Abstract

The use of the membrane probe fluorescein phosphatidylethanolamine (FPE) to investigate membrane binding is well established. However, until now, its use has been restricted to studies involving peptides and eukaryotic membranes. This useful tool has been developed to interrogate peptide:prokaryotic membrane interactions by introducing novel methodology to incorporate FPE into the membranes of UV killed, whole bacterial cells. The electrostatic potential of the membrane in the immediate vicinity of the probe affects the protonation state of the xanthene ring system in the fluorescein head group, which is held close to the membrane surface. When altered, e.g. by peptide binding and insertion, a change in fluorescence results, which can be measured spectrophotometrically. Applicability of this technique to bacterial surface interactions was confirmed by production of a binding curve for both a synthetic peptide and a 37kDa protein. Future investigations are anticipated to utilize this technology to characterize interactions of other toxins plus antimicrobial peptides such as lactoferricin and defensins with their target membranes.