# Structural Mechanism of Mammalian Prion Infectivity

> **NIH NIH R01** · DARTMOUTH COLLEGE · 2021 · $596,911

## Abstract

Project Summary
Mammalian prions, which cause fatal neurodegenerative diseases in humans and livestock
animals, are unorthodox infectious agents that replicate by the autocatalytic conformational
change of the host prion protein (PrPC) into a pathogenic isoform (PrPSc). Taking a reductionist
biochemical approach, we made the surprising discovery that endogenous cofactors such as
the membrane lipid phosphatidylethanolamine (PE) are required to produce infectious PrPSc
molecules. For example, autocatalytic PrPSc molecules formed with a combination of
recombinant PrP and PE substrates display a specific infectivity >105-fold greater than that of
similar autocatalytic PrPSc molecules formed from PrP alone. We have taken advantage of
this specific information to produce, for the first time, sufficient quantities of fully
infectious, isotopically labeled, and conformationally homogeneous recombinant prions
to perform high-resolution solid state (ss) NMR analysis of PrPSc structure. Here, our
collaborative team proposes to use ssNMR, electron microscopy, and mass spectrometry
analysis to elucidate the important structural elements that lead to infectivity. ssNMR will be
used to determine and compare the secondary structure maps of infectious and non-infectious
PrPSc molecules. This work will help identify and characterize specific infectivity-associated
domains as well as crucial NMR residue-peak assignments. Together with constraints from
electron microscopy and mass spectrometry analysis to identify the position of cut points using
digestion by proteinase K, these data will eventually enable structural determination of the entire
PrPSc molecule. We will also use ssNMR to determine precisely the structural mechanism by
which PE induces the infectious prion conformation. Finally, we will use 15N-13C transferred-
echo double resonance (TEDOR) to determine the symmetry pattern of PrPSc subunits in
isotopically mixed samples of infectious recombinant prions. These data will allow us to
discriminate between competing b-solenoid and in-register b-sheet quaternary structure models
of infectious PrPSc molecules. Overall, this proposal is a critical step towards determining
the full high-resolution structural determination of infectious mammalian prions as well
as the structural mechanism of prion infectivity.

## Key facts

- **NIH application ID:** 10191067
- **Project number:** 5R01NS118796-02
- **Recipient organization:** DARTMOUTH COLLEGE
- **Principal Investigator:** Surachai Supattapone
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $596,911
- **Award type:** 5
- **Project period:** 2020-06-15 → 2025-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10191067

## Citation

> US National Institutes of Health, RePORTER application 10191067, Structural Mechanism of Mammalian Prion Infectivity (5R01NS118796-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10191067. Licensed CC0.

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