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Research data in support of: Tailoring interleaflet lipid transfer with a DNA-based synthetic enzyme


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Authors

Ohmann, Alexander 
Joshi, Himanshu 
Aksimentiev, Aleksei 

Description

Data for "Tailoring interleaflet lipid transfer with a DNA-based synthetic enzyme" Nanoletters (2020). Further details can be found in the publication or its supporting information available free of charge at https://pubs.acs.org/doi/full/10.1021/acs.nanolett.0c00990

Files dithionite assays.zip contain micrographs of vesicles upon performing NBD reduction assay, from which time traces presented in Figure 3 of the paper (fluorescence vs time) were plotted. The images were collected on the Olympus confocal microscope, recorded every 10 s. FIJI was used to measure the pixel intensity, attributed to fluorescnce values. Files in current traces.zip contain transmembrane current traces for three published structures, as presented in Figure 2 (current vs time). These are ABF files collected with a pCLAMP software suite (using axopatch amplifier) upon applying 50 V across the DPhPC lipid membrane in the presence of bilayer-spanning DNA structures. LaserBleaching.zip contains datapoints collected during control experiment assessing laser bleaching (fluorescence vs time), presented in SI sup fig. 22. The experiment was performed in the same way (same microscope settings) as NBD reduction assay and presents marginal effects of laser bleaching during the said experiment. Two images gel_20_0 mM Mg.TIF and gel_20_1 mM Mg.PNG are the PAGE images obtained from a gel imaging setup, used for calculating the values presented in SI, sup fig. 15 and sup table 4. The bands correspond to: ladder (GeneRuler low range) and DNA duplex samples: 0D, 0fD, 1D, 2D, as described in detail in the associated publication. The gels were run at 100 V for 90 min, with DNA duplex samples folded at different concentrations of Mg. The strong effect of Mg on stability of the structures with internal modifications was deduced upon analysis. gel_20_0 mM Mg.TIF was run in the presence of 11 mM Mg in both the gel and the running buffer, while no Mg was present in the experiment resulting in gel_20_1 mM Mg.PNG. melingProfiles.xlsx is a file obtained from the spectrophotometer when collecting melting profiles for DNA structures (absorbance vs temperature) presented in SI, sup fig. 14, sup table 3. Absorbance at 260 nm of DNA duplexes at 1 uM concentration was collected in a 10-90 C range, with a rate of 1C/min.

Version

Software / Usage instructions

Origin, pCLAMP suite, FIJI

Keywords

DNA nanotechnology, lipid membrane, lipid scrambling, synthetic ion channel

Publisher

Sponsorship
European Research Council (647144)
EPSRC (1948702)
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