Abstract
Abstract
The elucidation of the atomistic structure of PrP
Sc
has, disappointingly, not revealed the mechanism underlying prion propagation. This is because PrP
Sc
has been shown to be a standard amyloid, and as such, it can only template a polypeptide chain that is unfolded. Therefore, while PrP
Sc
can easily template the disordered ∼90-120 domain of an incoming PrP
C
molecule, it does not have any obvious capacity to template its ∼121-231 folded domain (FD). The FD needs to accommodate into the ∼121-230 PrP
Sc
surface, an inert “Procrustean bed”. We have recently proposed that unfolding of the FD is a prerequisite for its conversion to the PrP
Sc
conformation and using experimental and computational methods identified the likely early events of such process using bank vole (BV)PrP(90-231) as our working model. We concluded that a key early event is the destabilization of the short β1-β2 assembly, and that the segment contiguous to the disordered tail, ∼121-140, encompassing β1 and its adjacent coils, is the most likely region to unfold first. In the present study we applied the same methodology to the disease-related PrP mutant E200K. E200K is the most prevalent mutation leading to familial prion disease in humans, yet the stability of its FD is only marginally lower compared to that of the protein with the wild type (wt) sequence. We, therefore, aimed at assessing whether the early unfolding events of BVPrP
C
(E200K) are similar or different to those of wtBVPrP
C
and whether such early unfolding route might provide insight into the extreme capacity of PrP
C
(E200K) to spontaneously generate PrP
Sc
. It should be noted that generation and propagation of PrP
Sc
are distinct phenomena, even though both are mechanistically related and dependent on conformational changes in PrP
C
. Our data show that early unfolding events of BVPrP
C
(E200K) are essentially similar to those of wt BVPrP
C
. However, interesting nuances exist.