Insertion of GPI-anchored alkaline phosphatase into supported membranes: a combined AFM and fluorescence microscopy study.
A new method based on combined atomic force microscopy (AFM) and fluorescence microscopy observations, is proposed to visualize the insertion of glycosylphosphatidyl inositol (GPI) anchored alkaline phosphatase from buffer solutions into supported phospholipid bilayers. The technique involves the use of 27 nm diameter fluorescent latex beads covalently coupled to the amine groups of proteins. Fluorescence microscopy allows the estimation of the relative protein coverage into the membrane and also introduces a height amplification for the detection of protein/bead complexes with the AFM. The coupling of the beads with the amine groups is not specific; this new and simple approach opens up new ways to investigate proteins into supported membrane systems.
Rieu
Jean-Paul
JP
Laboratoire de Physique de la Matière Condensée et Nanostructures, Université Claude Bernard Lyon-1, 69622 Villeurbanne, France. rieu@lpmcn.univ-lyon1.fr
Ronzon
Frédéric
F
Place
Christophe
C
Dekkiche
Fairouz
F
Cross
Benjamin
B
Roux
Bernard
B
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D006224:Cricetinae / Q000235:genetics; D017261:Glycosylphosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods
D017261:Glycosylphosphatidylinositols; D008051:Lipid Bilayers; D000469:Alkaline Phosphatase