title: Insertion of filamin into lipid membranes examined by calorimetry, the film balance technique, and lipid photolabeling.
abstract: The interaction of the actin-binding protein filamin with mixtures of zwitterionic and anionic phospholipids (DMPC, DMPG, PC, PS) was studied in reconstituted lipid monolayers and bilayers. Protein-lipid interactions were investigated by differential scanning calorimetry, the film balance technique, and hydrophobic photoradiolabeling. For calorimetric assays, multilamellar vesicles (MLVs) and large unilamellar vesicles produced by the extrusion technique (LUVETs) were used. With MLVs, filamin induced a pronounced drop in phase transition cooperativity. Mixed DMPC/DMPG LUVETs showed a linear decrease of the main phase transition enthalpy and a significant shift in temperature for the solidus and liquidus lines with increasing mole fractions of reconstituted filamin. The insertion of native filamin into uncharged and negatively charged lipid monolayers was measured in time/area diagrams with the film balance technique. Finally, we have newly synthesized a highly sensitive lipid analogue, [125]TID-PC/16, which selectively labels membrane-embedded hydrophobic domains of proteins, and which proved to label filamin, supporting evidence that this protein partially inserts into the hydrophobic domain of liposomes.
authors.affiliation: Department of Biophysics, Technical University of Munich, Garching, FRG.
authors.forename: M
authors.initials: M
authors.lastname: Tempel
authors.forename: W H
authors.initials: WH
authors.lastname: Goldmann
authors.forename: C
authors.initials: C
authors.lastname: Dietrich
authors.forename: V
authors.initials: V
authors.lastname: Niggli
authors.forename: T
authors.initials: T
authors.lastname: Weber
authors.forename: E
authors.initials: E
authors.lastname: Sackmann
authors.forename: G
authors.initials: G
authors.lastname: Isenberg
mesh_terms: D002152:Calorimetry, Differential Scanning; D003285:Contractile Proteins / Q000737:chemistry*; D064448:Filamins; D008563:Membrane Lipids / Q000737:chemistry*; D008567:Membranes, Artificial; D008840:Microfilament Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D010777:Photochemistry; D013816:Thermodynamics
chemical_list: D003285:Contractile Proteins; D064448:Filamins; D008563:Membrane Lipids; D008567:Membranes, Artificial; D008840:Microfilament Proteins