ABSTRACT
This review presents the recent advances in the achievement of organized proteo-lipidic nanostructures based on Langmuir-Blodgett technology and their potential applications in the nanobioscience area. By using the self-assembled properties of amphiphilic biomolecules at the air-water interface, the Langmuir-Blodgett (LB) technique offers the possibility to prepare ultrathin layers suitable for biomolecule immobilization at the molecular level. This review will provide a general overview of the enzyme association with preformed Langmuir-Blodgett films in connection with their potential applications in biosensing device developments, and then introduce the design of a new functionalised biomimetic nanostructure with oriented recognition site. The potential applications of such an organized proteo-lipidic nanostructure for biocatalysis investigations of an immobilised enzyme in a biomimetic situation and for the development of bioelectronic devices are finally discussed.
Subject(s)
Enzymes/chemistry , Lipids/chemistry , Membranes, Artificial , Nanotechnology/instrumentation , Nanotechnology/methods , Models, Biological , Surface PropertiesABSTRACT
This study deals with the kinetics properties of an enzyme immobilised in a defined orientation in a biomimetic environment. For this purpose, acetylcholinesterase (AChE) was captured at the surface of a nanostructured proteo-glycolipidic Langmuir-Blodgett film through specific recognition by a noninhibitor monoclonal antibody (IgG) inserted in a neoglycolipid bilayer. Modelling of this molecular assembly provided a plausible interpretation of the functional orientation of the enzyme. The AChE activity being stable for several weeks, the enzyme kinetics were investigated, and fitted perfectly with heterogeneous biocatalytic behaviour representative of cellular enzymatic catalysis. The AChE-IgG-glycolipid nanostructure was directly interfaced with an efficient optical device. Such an association, leading to an intimate contact between the nanostructure and the biochemical signal transducer, gives direct access to the intrinsic AChE behaviour. This study thus demonstrates the potential for direct investigation of the kinetic behaviour of an immobilised enzyme on a lipid bilayer through an efficient transduction system.
Subject(s)
Acetylcholinesterase/pharmacokinetics , Bungarus , Enzymes, Immobilized/pharmacokinetics , Lipid Bilayers , Snake Venoms/enzymology , Acetylcholinesterase/chemistry , Amino Acid Sequence , Animals , Enzyme Stability , Enzymes, Immobilized/chemistry , Luminescent Measurements , Models, Molecular , Molecular Sequence Data , Molecular Structure , Protein Conformation , Proteolipids/chemistry , Proteolipids/metabolism , Sequence Alignment , Snake Venoms/chemistryABSTRACT
The insertion of immunoglobulin (IgG) in a glycolipid monolayer was achieved by using the ability of new proteo-glycolipid vesicles to disintegrate into a mixed IgG-glycolipid interfacial film after spreading at an air-buffer interface. The interfacial disintegration kinetics was shown to be directly dependent on the initial vesicle surface density and on the buffer ionic strength. The presence of the immunoglobulin in the glycolipid film was displayed by an increase of the lateral compressibility (Cs) during monolayer compression. Cs magnitude modifications, due to the antibody effect on the monolayer packing, decreases as the spread vesicle density increases. At interfacial saturation, the lateral compressibility profile becomes similar to that of a control monolayer without antibody. However, the careful analysis of the mixed monolayer after transfer by Langmuir-Blodgett technique (ATR-FTIR characterisation, enzyme immunoassociation) clearly demonstrated that the antibody was still present in such conditions and was not completely squeezed out from the interface as compressibility changes could have meant. At nonsaturating vesicle surface density, IgG molecules initially lying in the lipid matrix with the Y-shape plane parallel to the interface move to a standing-up position during the compression, leading to lateral compressibility modifications. For a saturating vesicle surface density, the glycolipid molecules force the IgG molecules to directly adopt a more vertical position in the interfacial film and, consequently, no lateral compressibility modification was recorded during the compression.
