The potential role of copper binding sites in prion propagation

Transmissible spongiform encephalopathies (TSEs) or prion diseases are caused by a post-translational conversion of the normal cellular form of the prion protein (PrPC) into the pathological and infectious isoform denoted as prion or PrPSc. PrPC has been shown as a high-affinity copper-binding protein, and to a lesser extent binding to other divalent cations through the octarepeat region (OR) and the non-OR copper binding sites located in the disordered N-terminal domain. Studies on the role of copper in promoting prion conversion and infectivity yielded controversial results. In this work, we explored the role of histidine residues which are crucial for copper coordination in prion conversion using a combination of cell culture and cell-free approaches. The first evidence was derived from chronically prion-infected neuronal murine cells (ScN2a) transiently expressed in murine PrPC carrying artificial mutations at histidines located both at the OR and non-OR regions. We found that the lack of each histidine in the OR has neither effect on prion replication nor protein maturation and trafficking. Intriguingly, mutagenesis of histidine 95 (H95Y) does enhance prion conversion leading to de novo infectious material formation and cause aberrant accumulation during protein trafficking. Thus, we hypothesize that H95 could function as molecular switch for prion conversion, and copper bound to this residue may function in protein conformation stabilization. We also propose a cellular model for prion formation in cells expressing the H95Y mutant. Interestingly, our data may establish a platform for rationally designed experiments aimed at elucidating whether the H95Y mutation may cause de novo prion diseases in transgenic mice.