A DNA-Micropatterned Surface for Propagating Biomolecular Signals by Positional on-off Assembly of Catalytic Nanocompartments

Data underlying the figures in the publication: Maffeis, V. et al. “A DNA-Micropatterned Surface for Propagating Biomolecular Signals by Positional on-off Assembly of Catalytic Nanocompartments” Small 2022, 2202818, https://doi.org/10.1002/smll.202202818

Concept figures, Unicode origin graph (opj), TEM pictures, AFM pictures, LSM pictures

TOC

1. Figure 1 Concept figure representing the 3D view of micropatterned CNC immobilization promoting a cascade reaction between two distinct CNCs that ultimately results in a bioluminescent surface.
2. Figure 2 Schematic reactions involved in the biomolecular signal propagation.
3. Figure 3 Design and characterization of CNC-1 and CNC-2 (SLS and TEM).
4. Figure 4 Schematic rapresentation of DNA synthesis inside Klenow-CNCs by SYBR green I.
5. Figure 5 a) FCS autocorrelation curves of free Atto-488-template DNA (red) and Atto-488-template DNA CNC (blue); b) Activity of free Klenow polymerase, Klenow polymerase CNC with melittin, Klenow polymerase CNC without melittin at 25 °C; c) Enzyme activity of melittin-permeabilized Klenow CNCs and free Klenow fragment treated for 1 h at 55 °C; d) Enzyme activity of melittin-permeabilized Klenow CNCs and free Klenow fragment treated for 1 h at 75 °C.
6. Figure 6 a) Chemical functionalization of the micro-printed glass surface with the amino-functionalized ssDNA; b) AFM height image of the DNA-functionalized glass slide recorded in 10 mm Tris-HCl buffer at pH 7.2 at the resolution of 128 lines; c) CLSM image of glass surface microprinted with Cy5-labeled NH2-modified 31-mer.
7. Figure 7 a) left, AFM height image, middle, phase type image, and right, height profile (corresponding to dashed white line) of tandem CNCs attached via DNA hybridization on the microprinted glass surface recorded in 10 mm Tris-HCl buffer at pH 7.2; b) left, AFM height image, middle, phase type image and right, corresponding height profile of a single CNC; c) CLSM micrographs of polymersomes labeled with either cholesterol functionalized Atto-488 (green) or Dylight-633 (red)-DNA and immobilized by hybridization on a microprinted glass surface. Left panel, 488-channel, middle panel, 633-channel, right panel, merged image. Scale bars: 5 µm; d) Bioluminescence generation by permeable Klenow-CNCs and permeable ATP sulfurylase-CNCs (blue), by nonpermeable Klenow-CNCs and nonpermeable ATP sulfurylase-CNCs (pink), by the substrate mix alone (black), and by D-Luciferin and luciferase (red). Error bands represent ±SD, n = 3 replicates; e) QCM-D measurement of immobilized CNCs following repeated loading-removal cycles. Frequency (blue) and dissipation (brown) were recorded at three overtones (n = 3, 5, 7) as a function of time. (i, iv, vii) Addition of adaptor DNA, (ii, v, viii) immobilization of 22-mer polymersomes, and (iii, vi, ix) separation of DNA strands with 1 m NaOH.