Proteomics

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Altered Domain Structure of the Prion Protein Caused by Cu2+ Binding and Functionally Relevant Mutations: Analysis by Cross‐Linking, MS/MS, and NMR


ABSTRACT: The endogenous cellular prion protein (PrPC) can misfold into the scrapie isoform (PrPSc) and cause fatal infectious diseases. Despite significant research on the prion protein, both its normal function and whether alterations to that function play a critical role in prion diseases remain unknown. The protein consists of a predominantly alpha-helical C-terminal domain and an unstructured N-terminal domain that can coordinate Cu2+. Previous studies using NMR and EPR have revealed a tertiary association between the N-terminal domain and the C-terminal domain that we have hypothesized to be critical to the protein’s normal function. Here we investigated and quantified the inter-domain interactions within three different prion variants (wild type recombinant mouse PrPC, mutant delta central region (ΔCR), and disease mutant (E199K) after chemical cross-linking with a newly designed MS-cleavable reagent 1-(4-((2,5-Dioxopyrrolidin-1-yl)oxy)-4-oxobutyl)-4-(2-(3-methyl-3H-diazirin-3-yl)ethyl)-1,4-diazabicyclo[2.2.2] octane-1,4-diium (APDC4), followed by nHPLC(RP) and tandem MS analysis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Catherine Costello  

LAB HEAD: Catherine E. Costello

PROVIDER: PXD012427 | Pride | 2020-06-16

REPOSITORIES: Pride

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Publications

Altered Domain Structure of the Prion Protein Caused by Cu<sup>2+</sup> Binding and Functionally Relevant Mutations: Analysis by Cross-Linking, MS/MS, and NMR.

McDonald Alex J AJ   Leon Deborah R DR   Markham Kathleen A KA   Wu Bei B   Heckendorf Christian F CF   Schilling Kevin K   Showalter Hollis D HD   Andrews Philip C PC   McComb Mark E ME   Pushie M Jake MJ   Costello Catherine E CE   Millhauser Glenn L GL   Harris David A DA  

Structure (London, England : 1993) 20190404 6


The cellular isoform of the prion protein (PrP<sup>C</sup>) serves as precursor to the infectious isoform (PrP<sup>Sc</sup>), and as a cell-surface receptor, which binds misfolded protein oligomers as well as physiological ligands such as Cu<sup>2+</sup> ions. PrP<sup>C</sup> 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 spe  ...[more]

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