Abstract
According to the current model of nerve propagation, the function of acetylcholinesterase (AChE) is to terminate synaptic transmission of nerve signals by hydrolyzing the neurotransmitter acetylcholine (ACh) in the synaptic cleft to acetic acid (acetate) and choline. However, extra-synaptic roles, which are known as ‘non-classical’ roles, have not been fully elucidated. Here, we measured AChE activity with the enzyme bound to lipid membranes of varying area per enzyme in vitro using the Ellman assay. We found that the activity was not affected by density fluctuations in a supported lipid bilayer (SLB) induced by standing surface acoustic waves. Nevertheless, we found twice as high activity in the presence of small unilamellar vesicles (SUV) compared to lipid-free samples. We also showed that the increase in activity scaled with the available membrane area per enzyme.
Highlights
The lateral organization of the cell membrane, including so-called lipid rafts, is essential for the formation of functional units in biology [1,2,3,4]
A small amount of fluorescent dye molecules is added to the supported lipid bilayers (SLB)
We investigated the binding efficiency of the enzyme to the SLB as well as the influence of a Surface acoustic waves (SAW) standing wave field on the activity of the bound enzyme
Summary
The lateral organization of the cell membrane, including so-called lipid rafts, is essential for the formation of functional units in biology [1,2,3,4]. Hochrein et al studied the conformational behaviour of DNA molecules adsorbed on cationic lipid membranes deposited on grooved, onedimensional, periodic, microstructured surfaces [9]. Another patterning technique uses proteins and PDMS stamps to induce patterns in SLB [10]. This apparent shortcoming was pointed out by Jacobson et al.: “the field of lipid rafts is currently at a technical impasse, as the physical tools to study biological membranes as spatially and temporally ordered fluid are still being developed.” [4] Surface acoustic waves (SAW) with amplitudes of the order of 1 nm and variable wavelength and frequency between about 30 μm at 100 MHz and 3 μm at 1000 MHz can be used to generate standing waves and a tunable energy landscape
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