[en] Atomic force microscopy (AFM)-based single-molecule force spectroscopy is a prevalent tool for the exploration of individual (bio)molecules, providing exquisite information on many molecular-level processes. For example, proteins, DNA, polysaccharides, supramolecular polymers and polyelectrolytes have been investigated, revealing details about the strength of intramolecular interactions, folding and unfolding pathways, mechanics, conformational changes, reactivity, kinetics, etc.
For each particular system under investigation, the experimental design is a decisive phase that often involves a multistep chemical protocol, including grafting, derivatization, coupling, (de-)protection, and other functionalization reactions. Procedures of sample preparation are often complex and time-consuming. Hence, there is a need for new general platforms allowing for straightforward sample preparation adapted to single-molecule studies, i.e. a tight attachment to both the substrate and the tip, and a low density to favor single-molecule detection.
We report here on the use of poly(ethylene oxide) (PEO) as a tether to probe various properties of individual molecules. The polymeric linker acts as a handle that stably binds to the AFM tip. The simple adsorption of poly(ethylene oxide) to the tip is versatile and provides an appropriate system configuration for the investigation of many different biological and synthetic molecular systems. To attest for this versatility and adequacy with advanced single-molecule investigation, we present different examples of PEO-mediated studies about the unfolding of a synthetic peptide, the mechanochemical behavior of a molecular machine and finally the stability of a metallo-supramolecular complexed polymer.
All the requirements for the study of peptide conformation, tiny molecular machines or metallo-supramolecular interactions in solution are here fulfilled. More generally, this method based on non-covalent sorption of PEO on an AFM tip, can be implemented in a wide range of solvents, for the study of many intra- or intermolecular phenomena at the single-molecule level over orders of magnitude of force loading rates. Connecting PEO tethers to a very broad variety of (bio)molecules is a facile and versatile route. The commercial availability of many different functional PEOs makes this tethering strategy even more accessible.
