1. Altered Domain Structure of the Prion Protein Caused by Cu 2+ Binding and Functionally Relevant Mutations: Analysis by Cross-Linking, MS/MS, and NMR.
- Author
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McDonald AJ, Leon DR, Markham KA, Wu B, Heckendorf CF, Schilling K, Showalter HD, Andrews PC, McComb ME, Pushie MJ, Costello CE, Millhauser GL, and Harris DA
- Subjects
- Animals, Cell Line, Copper metabolism, Cross-Linking Reagents chemistry, Cross-Linking Reagents metabolism, Humans, Magnetic Resonance Spectroscopy methods, Mice, Prion Proteins metabolism, Protein Binding, Protein Isoforms chemistry, Protein Isoforms genetics, Protein Isoforms metabolism, Tandem Mass Spectrometry methods, Copper chemistry, Molecular Dynamics Simulation, Mutation, Prion Proteins chemistry, Prion Proteins genetics, Protein Domains
- Abstract
The cellular isoform of the prion protein (PrP
C ) serves as precursor to the infectious isoform (PrPSc ), and as a cell-surface receptor, which binds misfolded protein oligomers as well as physiological ligands such as Cu2+ ions. PrPC consists of two domains: a flexible N-terminal domain and a structured C-terminal domain. Both the physiological and pathological functions of PrP depend on intramolecular interactions between these two domains, but the specific amino acid residues involved have proven challenging to define. Here, we employ a combination of chemical cross-linking, mass spectrometry, NMR, molecular dynamics simulations, and functional assays to identify residue-level contacts between the N- and C-terminal domains of PrPC . We also determine how these interdomain contacts are altered by binding of Cu2+ ions and by functionally relevant mutations. Our results provide a structural basis for interpreting both the normal and toxic activities of PrP., (Copyright © 2019 Elsevier Ltd. All rights reserved.)- Published
- 2019
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