Post-translational modifications in PrP expand the conformational diversity of prions in vivo

Patricia Aguilar-Calvo,Hasier Eraña,Joaquin Castilla,Xiangzhu Xiao,K Peter R Nilsson,Christina J Sigurdson,Witold K Surewicz,Cyrus Bett,Katrin Soldau

doi:10.1038/srep43295

Abstract

Misfolded prion protein aggregates (PrPSc) show remarkable structural diversity and are associated with highly variable disease phenotypes. Similarly, other proteins, including amyloid-β, tau, α-synuclein, and serum amyloid A, misfold into distinct conformers linked to different clinical diseases through poorly understood mechanisms. Here we use mice expressing glycophosphatidylinositol (GPI)-anchorless prion protein, PrPC, together with hydrogen-deuterium exchange coupled with mass spectrometry (HXMS) and a battery of biochemical and biophysical tools to investigate how post-translational modifications impact the aggregated prion protein properties and disease phenotype. Four GPI-anchorless prion strains caused a nearly identical clinical and pathological disease phenotype, yet maintained their structural diversity in the anchorless state. HXMS studies revealed that GPI-anchorless PrPSc is characterized by substantially higher protection against hydrogen/deuterium exchange in the C-terminal region near the N-glycan sites, suggesting this region had become more ordered in the anchorless state. For one strain, passage of GPI-anchorless prions into wild type mice led to the emergence of a novel strain with a unique biochemical and phenotypic signature. For the new strain, histidine hydrogen-deuterium mass spectrometry revealed altered packing arrangements of β-sheets that encompass residues 139 and 186 of PrPSc. These findings show how variation in post-translational modifications may explain the emergence of new protein conformations in vivo and also provide a basis for understanding how the misfolded protein structure impacts the disease.

Highlights

Misfolded prion protein aggregates (PrPSc) show remarkable structural diversity and are associated with highly variable disease phenotypes
We previously found that passage of one anchorless prion strain (GPI-RML, second passage) into WT mice may have led to the reemergence of the original RML strain[32]
To better understand how post-translational modifications (PTMs) impact the conformation of protein aggregates and how PTMs underlie the generation of new strains in vivo, here we investigate the fidelity of diverse prions during transit into a GPI-anchorless state and back into a GPI-anchored state

Summary

Introduction

Misfolded prion protein aggregates (PrPSc) show remarkable structural diversity and are associated with highly variable disease phenotypes. We use mice expressing glycophosphatidylinositol (GPI)anchorless prion protein, PrPC, together with hydrogen-deuterium exchange coupled with mass spectrometry (HXMS) and a battery of biochemical and biophysical tools to investigate how posttranslational modifications impact the aggregated prion protein properties and disease phenotype. Four GPI-anchorless prion strains caused a nearly identical clinical and pathological disease phenotype, yet maintained their structural diversity in the anchorless state. Passage of GPI-anchorless prions into wild type mice led to the emergence of a novel strain with a unique biochemical and phenotypic signature. The origin of these diverse protein conformational subtypes that emerge within an individual or within a population, and how these markedly different structures impact the disease outcome, is unclear. We previously found that passage of one anchorless prion strain (GPI-RML, second passage) into WT mice may have led to the reemergence of the original RML strain[32]

Methods

Results

Conclusion