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Surface plasmon resonance (SPR) employs the optical principle of SPR to measure changes in mass on a sensor chip surface in real time. Surface chemistry has been developed which enables the immoblization of lipid bilayers and determination of protein-membrane interactions in real time. In the last decade, the plasma membrane has been demonstrated to play an important role in amyloidogenesis and cytotoxicity induced by amyloidogenic proteins. SPR provides an ideal way to study the membrane binding of amyloidogenic proteins. In this chapter, we describe the application of SPR to the study of amyloidogenic transthyretin binding to the plasma membrane and artificial lipid bilayers.

Surface plasmon resonance (SPR) employs the optical principle of SPR to measure changes in mass on a sensor chip surface in real time. Surface chemistry has been developed which enables the immobilization of lipid bilayers and determination of protein-membrane interactions in real time. Antimicrobial peptides are being increasingly recognized as potential candidate antibacterial drugs in the face of the rapidly emerging bacterial resistance to conventional antibiotics in recent years. However, a precise understanding of the relationship between antimicrobial peptide structure and their cytolytic function in a range of organisms is still lacking. This is a result of the complex nature of the interactions of antimicrobial peptides with the cell membrane, the mechanism of which can vary considerably between different classes of antimicrobial peptides. SPR has recently been applied to the study of biomembrane-based systems which has allowed a real-time analysis of binding affinity and kinetics. This chapter describes an SPR method to study the membrane interactions of melittin, a well-known antimicrobial peptide.

Surface plasmon resonance (SPR) spectroscopy is emerging as a useful tool for determination of molecular interactions in real time. Studies on the molecular pathogenesis of amyloidoses have shown that the plasma membrane plays an important role in amyloidogenesis and cytotoxicity induced by amyloidogenic proteins. By immobilizing lipid bilayers on a sensor chip surface, SPR spectroscopy has been employed to examine the binding of amyloidogenic proteins, such as amyloid beta protein (Abeta), to a variety of lipid membranes, and it provided new insights into the molecular interactions between these amyloidogenic proteins and membranes. In this chapter, we describe the application of SPR spectroscopy to the determination of the binding of Abeta to lipid membranes.

The chromatographic support containing monolayers of phospholipids offers novel modes in analyzing and separating proteins. The polar choline head groups on immobilized phosphatidylcholine were used for the affinity purification of phospholipase A (PLA). The purification process involves removing the contaminating proteins with detergent additives to the elution buffer such as short-chain alkylsulfonates. The lipid-bound PLA was eluted with acetonitrile or octyllysophosphatidylcholine. The purity of PLA was approximately 70% based on densitometric scans of gel electrophoresis. These results suggest that the lipid-immobilized chromatography may be applied to develop purification methods for PLA, enzymes, and membrane proteins obtained from diverse cells.

The use of specific quenched fluorescent substrates (QFS) provides a rapid and sensitive method to measure peptidase activity, and is readily adaptable to high-throughput screening of potential peptidase inhibitors. In this chapter, we discuss general considerations for the development of QFS assays, and describe in detail an assay protocol for the mammalian metallopeptidase, endothelin-converting enzyme.