Application of different lipid surfaces to monitor protein–membrane interactions by surface plasmon resonance spectroscopy

Article type: Research Article

Authors: Fischer, Torsten^; | Senin, Ivan I. | Philippov, Pavel P. | Koch, Karl‐Wilhelm^;

Affiliations: Institut für Biologische Informationsverarbeitung 1, Forschungszentrum Jülich, D‐52425 Jülich, Germany | A.N. Belozersky Institute of Physico‐Chemical Biology, Moscow State University, 119992 Moscow, Russia

Note: [] Current address: Institute of Genetic Medicine and Neurogenetic Institute, Departament of Biochemistry and Molecular Biology, UCLA, CA 90033, USA.

Note: [] Corresponding author. Tel.: +49 2461 61 3255; Fax: +49 2461 614216; E‐mail: k.w.koch@fz‐juelich.de.

Abstract: Planar lipid bilayers on sensor chip surfaces have become useful tools to study membrane bound processes by surface plasmon resonance spectroscopy. We immobilized phospholipids on sensor chips by different approaches. First, a self‐assembled monolayer of octadecylmercaptan was formed on a blank gold surface and subsequent addition of phospholipids led to formation of a heterobilayer. Second, a self‐assembled monolayer of mercaptoundecanoic acid was formed on a gold surface, the carboxy groups of mercaptoundecanoic acid were activated and covalently linked to phosphatidylethanolamine. Addition of phospholipids then led to a bilayer with phosphatidylethanolamine as the lower leaflet. Third, a hydrophobic sensor chip (L1, BIAcore) was used as a binding matrix for phospholipids. These lipid surfaces were tested, whether they are suitable to study protein–membrane interactions. As biological test system we used the Ca2+‐myristoyl‐switch of the neuronal Ca2+‐binding protein recoverin. All three surfaces were sufficiently stable to monitor the Ca2+‐dependent binding of recoverin to membranes.

Journal: Spectroscopy, vol. 16, no. 3-4, pp. 271-279, 2002