title: Importance of phospholipid bilayer integrity in the analysis of protein-lipid interactions.
abstract: The integrity of supported phospholipid bilayer membranes is of crucial importance for the investigation of lipid-protein interactions. Therefore we recorded the formation of supported membranes on SiO2 and mica by quartz crystal microbalance and controlled the integrity by atomic force microscopy. This study aims to analyze how membrane defects affect protein-lipid interactions. The experiments focused on a lipid mixture of POPC/DOPC/Chol/POPS/PI(4,5)P2 (37:20:20:20:3) and the binding of the peripheral membrane associated protein annexin A2. We found that formation of a continuous undisturbed bilayer is an indispensable precondition for a reliable determination and quantification of lipid-protein-interactions. If membrane defects were present, protein adsorption causes membrane disruption and lipid detachment on a support thus leading to false determination of binding constants. Our results obtained for PI(4,5)P2 and cholesterol containing supported membranes yield new knowledge to construct functional surfaces that may cover nanoporous substrates, form free standing membranes or may be used for lab-on-a-chip applications.
authors.affiliation: Institute of Biochemistry, University of Münster, Wilhelm-Klemm-Str. 2, D-48149 Münster, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Drücker
authors.affiliation: Institute of Medical Biochemistry, ZMBE, University of Münster, Von-Esmarch-Str. 56, D-48149 Münster, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Gerke
authors.affiliation: Institute of Biochemistry, University of Münster, Wilhelm-Klemm-Str. 2, D-48149 Münster, Germany. Electronic address: gallah@uni-muenster.de.
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Galla
mesh_terms: D000327:Adsorption; D000538:Aluminum Silicates; D017306:Annexin A2 / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D058525:Quartz Crystal Microbalance Techniques; D012822:Silicon Dioxide; D013499:Surface Properties
chemical_list: D000538:Aluminum Silicates; D017306:Annexin A2; D008051:Lipid Bilayers; D010743:Phospholipids; D012822:Silicon Dioxide; C011934:mica
keywords: Cholesterol; Lipid protein interactions; Phosphatidylinositol-(4,5)-bisphosphate (PIP(2), PI(4,5)P(2)); Solid supported bilayer