Controlled enhancement of transmembrane enzyme activity in polymer cushioned supported bilayer membranes
- 7 September 2010
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Soft Matter
- Vol. 6 (21), 5382-5389
- https://doi.org/10.1039/c0sm00565g
Abstract
Reconstitution of transmembrane proteins into supported lipid bilayer (SLB) membranes has often been hampered by strong interactions of protein domains with the underlying solid support leading to loss of both activity and mobility within the plane of the lipid bilayer. Polymer cushioned SLBs can overcome this by avoiding direct contact with the support. To extend this approach we developed an anionic polymer cushion system that allows tunable lipid mobility as well as functional integration of the transmembrane protein β-amyloid precursor protein cleaving enzyme (BACE) into SLBs. Fluorescence recovery after photobleaching analysis revealed a homogeneous distribution and high lateral mobility of the reconstituted BACE in cushioned SLBs while an impaired mobility and inhomogeneous clustering of reconstituted BACE were found in SLBs on silicon oxide substrates. The cushioning of SLBs led to increased incorporation and enhanced enzymatic activity of the reconstituted BACE with a direct correlation between lipid mobility and BACE activity. The utilized polymer cushion system allows the successful reconstitution of transmembrane proteins within SLBs with tunable properties.Keywords
This publication has 33 references indexed in Scilit:
- Supported Lipid Bilayers on Spacious and pH-Responsive Polymer Cushions with Varied HydrophilicityThe Journal of Physical Chemistry B, 2008
- Insulating tethered bilayer lipid membranes to study membrane proteinsMolecular BioSystems, 2007
- Reversible Activation of Diblock Copolymer Monolayers at the Interface by pH Modulation, 2: Membrane Interactions at the Solid/Liquid InterfaceThe Journal of Physical Chemistry B, 2006
- Lipids as Modulators of Proteolytic Activity of BACEPublished by Elsevier BV ,2005
- Differences between non-specific and bio-specific, and between equilibrium and non-equilibrium, interactions in biological systemsQuarterly Reviews of Biophysics, 2005
- Cell biology of protein misfolding: The examples of Alzheimer's and Parkinson's diseasesNature, 2004
- In a mirror dimly: tracing the movements of molecules in living cellsTrends in Cell Biology, 2004
- Folding proteins in fatal waysNature, 2003
- Simultaneous in Situ Total Internal Reflectance Fluorescence/Atomic Force Microscopy Studies of DPPC/dPOPC Microdomains in Supported Planar Lipid BilayersJournal of the American Chemical Society, 2003
- Production of large unilamellar vesicles by a rapid extrusion procedure. Characterization of size distribution, trapped volume and ability to maintain a membrane potentialBiochimica et Biophysica Acta (BBA) - Biomembranes, 1985