S. Roger Qiu

A multistage pathway for human prion protein aggregation in vitro: from multimeric seeds to β-oligomers and nonfibrillar structures.

Aberrant protein aggregation causes numerous neurological diseases including Creutzfeldt-Jakob disease (CJD), but the aggregation mechanisms remain poorly understood. Here, we report AFM results on the formation pathways of β-oligomers and nonfibrillar aggregates from wild-type full-length recombinant human prion protein (WT) and an insertion mutant (10OR) with five additional octapeptide repeats linked to familial CJD. Upon partial denaturing, seeds consisting of 3-4 monomers quickly appeared. Oligomers of ~11-22 monomers then formed through direct interaction of seeds, rather than by subsequent monomer attachment. All larger aggregates formed through association of these β-oligomers. Although both WT and 10OR exhibited identical aggregation mechanisms, the latter oligomerized faster due to lower solubility and, hence, thermodynamic stability. This novel aggregation pathway has implications for prion diseases as well as others caused by protein aggregation.

Growth inhibition of calcium oxalate monohydrate crystal by linear aspartic acid enantiomers investigated by in situ atomic force microscopy

The inhibitory effect of linear enantiomers of L- and D-Asp6 on the growth of calcium oxalate monohydrate crystal has been investigated using in situ atomic force microscopy. The inhibitory magnitude of D-Asp6 on the growth of the [00] step on the (010) face is about 10% larger than that of L-Asp6. While no chiral effect is observed or expected on the growth of the [0] step on the (01) face by both enantiomers, their inhibitory effect on this step is much stronger than that on the [00] step on the (010) face. In both cases, the step morphology indicates that these enantiomers create the impurity pinning along the steps, while the dependence of step speed on supersaturation shows that they also produce a reduction of the step kinetic coefficients. Analysis of the step speed data within the context of an existing model for step pinning and kink blocking shows that the major impact of Asp6 is to block active kink sites. The larger inhibition of the [00] step growth by D-Asp6 over L-Asp6 and the substantially larger inhibition of the [0] step over the [00] step by both enantiomers both result from larger affinity for adsorption to the (010) face and the (01) face, respectively. This is because the larger adsorption leads to a higher density of blocking kink sites along the steps. The estimated difference in binding energy of L- and D-Asp6 to the respective faces from the kinetics model is consistent with the trend predicted by our molecular modeling of the enantiomer binding to the faces.