Insertion of GPI-anchored alkaline phosphatase into supported membranes: a combined AFM and fluorescence microscopy study.

• Authors: Jean-Paul Rieu , Frédéric Ronzon , Christophe Place , Fairouz Dekkiche , Benjamin Cross , Bernard Roux
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

fluorescent beads; atomic force microscopy (AFM); GPI anchors; alkaline phosphatase; supported lipid membranes

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2004
• Volume: 51
• Issue: 1
• Pages: 189-197

Dates

published

2004

received

2003-12-19

revised

2004-01-07

accepted

2004-02-06

Contributors

author

Jean-Paul Rieu

• Laboratoire de Physique de la Matière Condensée et Nanostructures, Université Claude Bernard Lyon-1, 69622 Villeurbanne, France

author

Frédéric Ronzon

• Laboratoire de Physique de la Matière Condensée et Nanostructures, Université Claude Bernard Lyon-1, 69622 Villeurbanne, France

author

Christophe Place

• Laboratoire de Physique, ENS Lyon, 69364 Lyon cedex 07, Lyon, France

author

Fairouz Dekkiche

• Laboratoire Pollution et Traitement des Eaux, Université Mentouri-Constantine, 25000 Constantine, Algeria

author

Benjamin Cross

• Laboratoire de Physique de la Matière Condensée et Nanostructures, Université Claude Bernard Lyon-1, 69622 Villeurbanne, France

author

Bernard Roux

• Laboratoire de Physique de la Matière Condensée et Nanostructures, Université Claude Bernard Lyon-1, 69622 Villeurbanne, France

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