Model to rationalize and predict the formation of organic patterns originating from an enzyme-assisted self-assembly Liesegang-like process of peptides in a host hydrogel.

Recently, we have investigated the enzyme-assisted self-assembly of precursor peptides diffusing in an enzyme-containing host gel, leading to various self-assembly profiles within the gel. At high enzyme concentrations, the reaction-diffusion self-assembly processes result in the formation of a continuous non-monotonous peptide self-assembly profile. At low enzyme concentrations, they result in the formation of individual self-assembled peptide microglobules and at intermediate enzyme concentrations both kinds of self-assembled structures coexist. Herein, we develop a Liesegang-type model that considers four major points: (i) the diffusion of the precursor peptides within the host gel, (ii) the diffusion of the enzymes in the gel, (iii) the enzymatic transformation of the precursor peptides into the self-assembling ones and (iv) the nucleation of these building blocks as the starting point of the self-assembly process. This process is treated stochastically. Our model predicts most of the experimentally observed features and in particular (i) the transition from a continuous to a microglobular pattern of self-assembled peptides through five types of patterns by decreasing the enzyme concentration in the host hydrogel. (ii) It also predicts that when the precursor peptide concentration decreases, the enzyme concentration at which the continuous/microglobules transition appears increases. (iii) Finally, it predicts that for peptides whose critical self-assembly concentration in solution decreases, the peptide concentration at which the continuous-to-microglobular transition decreases too. All these predictions are observed experimentally.

Reversible Modulation of Aptamer-Ligand Binding in RNA Light-Up Aptamers Containing G-Quadruplex Using Chemical Stimuli.

Regulating a system in equilibrium transiently to out-of-equilibrium by using certain stimuli is the strategy used by natural biomolecules to function. Herein, we showed that the interaction of synthetic RNA aptamers, having a G-quadruplex core structure, with their corresponding ligands could be regulated from their equilibrium state to non-equilibrium state in a reversible manner using simple chemical stimuli (Ag+ and cysteine). The approach would be useful for designing aptamer regulators that work in a dynamic nucleic acid network, where a strict control on aptamer-ligand interaction is needed. In addition, to the best of our knowledge, this is the first report which shows that RNA G-quadruplexes can be disrupted by the addition of silver ions. This would be useful not only in designing RNA-based sensors or regulators but would also be useful for understanding the role of metal ions in RNA folding and catalysis.

Conformation and Morphology of 4-(NH2/OH)-Substituted l/d-Prolyl Polypeptides: Effect of Homo- and Heterochiral Backbones on Formation of ß-Structures and Nanofibers.

The relative stereochemistry of C2 and C4 in 4-substituted prolyl polypeptides plays an important role in defining the derived conformation in solution. cis-(2S,4S)-Amino/hydroxy-l-prolyl polypeptide (lC-Amp 9/lC-Hyp 9) shows a PPII conformation in phosphate buffer and a ß-structure in a relatively hydrophobic solvent, trifluoroethanol (TFE). It is now demonstrated that the homochiral enantiomeric cis-substituted d-prolyl polypeptide (dC-Amp 9/dC-Hyp 9) exhibits mirror image ß-structures in TFE. In the case of alternating heterochiral prolyl peptides, it is the trans-substituted [lT(2S,4R)-dT(2R,4S)] n prolyl polypeptide that shows ß-structures in TFE, while the cis-substituted [lC(2S,4S)-dC(2R,4R)] n prolyl polypeptide is disordered in both phosphate buffer and TFE. The results highlight the important chirality-specific structural requirements for ß-structure formation. The observed conformation in solution (circular dichroism (CD)) is also correlated with the morphology of the self-assemblies (field emission scanning electron microscopy (FESEM)), with the PPII form leading to spherical nanoparticles and ß-structures leading to nanofiber formation. The results shed light on the role of relative stereochemistry at C2 and C4 in defining the polyproline peptide conformation in solution and how different conformations drive self-assemblies of peptides toward specific nanostructures.