No abstract

We report on a new highly reproducible route to recognitive self-assembly of molecular-scale circuits using sulfur-terminated subnanometer diameter Mo6S9-xIx (MoSIx) molecular nanowires. We demonstrate solution-processed attachment of MoSIx connecting leads to gold nanoparticles (GNPs). We also show that naked nanowires have the potential to bind thiolated proteins such as green fluorescent protein directly, thus providing a universal construct to which almost any protein could be attached. We further demonstrate three-terminal branched circuits with GNPs, opening a self-assembly route to multiscale complex molecular-scale architectures at the single-molecule level.

Amyloid-induced toxicity is a well-known phenomenon but the molecular background remains unclear. One hypothesis relates toxicity to amyloid-membrane interactions, predicting that amyloid oligomers make pores into membranes. Therefore, the toxicity and membrane interaction of prefibrillar aggregates and individual oligomers of a non-pathological yet highly amyloidogenic protein human stefin B (cystatin B) was examined. By monitoring caspase-3 activity and by testing cell viability, we showed that the lag phase aggregates obtained at pH 5 and 3 were toxic to neuroblastoma cells. Of equal toxicity were the higher-order oligomers prepared at pH 7 by freeze-thaw cycles. The higher-order oligomers eluted on size-exclusion chromatography (SEC) as a broad peak comprising hexamers, octamers, 12- and 16-mers, well separated from monomers, dimers and tetramers. Only oligomers higher than the tetramers (Rh >3.5 nm) proved toxic, in contrast to dimers and tetramers. In accordance with data from SEC, dynamic light scattering and atomic force microscopy data indicate that the toxic oligomers have diameters larger than 4 nm. Critical pressure measurements showed that the toxic higher-order oligomers inserted more effectively into model lipid monolayers than dimers and tetramers. They also bound, similarly to prefibrillar aggregates, to the plasma membrane and became internalized. Taken together, our results confirm the importance of membrane interaction and perforation in the phenomenon of cytotoxicity.

It is well established that contact order and folding rates are correlated for small proteins. The folding rates of stefins A and B differ by nearly two orders of magnitude despite sharing an identical native fold and hence contact order. We break down the determinants of this behavior and demonstrate that the modulation of contact order effects can be accounted for by the combined contributions of a framework-like mechanism, characterized by intrinsic helix stabilities, together with nonnative helical backbone conformation and nonnative hydrophobic interactions within the folding transition state. These contributions result in the formation of nonnative interactions in the transition state as evidenced by the opposing effects on folding rate and stability of these proteins.

We report a simple method of functionalization of MoSIx nanowires with biomaterials via the thiol group. We succeed to immobilize iodinated and sulphated high molecular mass thyroglobulin proteins on MoSIx nanowires. By controlling the preparation of the nanowires before self-assembly we can attach proteins either at the ends or achieve complete wrapping of the nanowires. The control parameter is the S and I stoichiometry ratio of the MoSIx nanowires. Breaking force measurements were performed in order to confirm and characterise the wrapping of the nanowires with the proteins.