Fig. 2: Ribozyme activity within coacervates dispersions.
From: RNA-peptide interactions tune the ribozyme activity within coacervate microdroplet dispersions
A Schematic showing the experimental design to measure HHmin reaction kinetics inside coacervate dispersions. 250 µM HHmin was mixed with 500 µM peptides to generate coacervate droplets. 50 µM FAM-tagged substrate was gently added to the dispersion. After each time point, the mixture was centrifuged, and 7 µL of the supernatant was removed. The pellet containing coacervate was then analysed by 20% urea-PAGE to visualise the cleaved and uncleaved FAM-substrate separated by gel electrophoresis. B Representative gel electrophoresis image showing comparative substrate and product band fluorescence intensities after 6 h. HHmin was mixed with 10% FAM-tagged HHmin to see the relative ribozyme concentration inside each system. Ci Plots showing product formation (%) as a function of time for each of the HHmin/peptide coacervate systems. Circles are the data points, and the dotted line shows a representative first-order reaction kinetics model fitted to the averaged data points. Error bars indicate the standard deviation from three experimental replicates. Cii Apparent rate constant (k) comparison between six HHmin/peptide coacervate systems. The apparent rate constants were obtained from fitting the product formed as a function of time, using first-order reaction kinetics. The bar plot shows the average apparent rate constants from at least three independent experiments and the error bars show the standard deviation. Source data is provided in the repository.
