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Fungal prion HET-s as a model for structural complexity and self-propagation in prions.

Proceedings of the National Academy of Sciences of the United States of America (2014-04-08)
William Wan, Gerald Stubbs
ABSTRACT

The highly ordered and reproducible structure of the fungal prion HET-s makes it an excellent model system for studying the inherent properties of prions, self-propagating infectious proteins that have been implicated in a number of fatal diseases. In particular, the HET-s prion-forming domain readily folds into a relatively complex two-rung β-solenoid amyloid. The faithful self-propagation of this fold involves a diverse array of inter- and intramolecular structural features. These features include a long flexible loop connecting the two rungs, buried polar residues, salt bridges, and asparagine ladders. We have used site-directed mutagenesis and X-ray fiber diffraction to probe the relative importance of these features for the formation of β-solenoid structure, as well as the cumulative effects of multiple mutations. Using fibrillization kinetics and chemical stability assays, we have determined the biophysical effects of our mutations on the assembly and stability of the prion-forming domain. We have found that a diversity of structural features provides a level of redundancy that allows robust folding and stability even in the face of significant sequence alterations and suboptimal environmental conditions. Our findings provide fundamental insights into the structural interactions necessary for self-propagation. Propagation of prion structure seems to require an obligatory level of complexity that may not be reproducible in short peptide models.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Asparagine, ≥98% (HPLC)
Sigma-Aldrich
L-Asparagine, BioReagent, suitable for cell culture, suitable for insect cell culture
Supelco
L-Asparagine, certified reference material, TraceCERT®