Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation

Kaneko, Kiyotoshi; Zulianello, Laurence; Scott, Michael; Cooper, Carol M.; Wallace, Andrew; James, Thomas Leroy; Cohen, Fred E.; Prusiner, Stanley B.

doi:10.1073/pnas.94.19.10069

Cited by 469 publications

(447 citation statements)

References 44 publications

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“…Intriguingly, residues Q167, Q171, V214, and Q218 have been identified as the binding site for the socalled ''protein X''. 56 These residues exhibit, or are sequentially adjacent to residues that exhibit, R ex motions. The binding of an as yet, unidentified molecule might destabilize PrP C or stabilize a PrP* intermediate.…”

Section: Prp C Folding Intermediates and Stabilitymentioning

confidence: 99%

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

et al. 2009

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Prion diseases are associated with the misfolding of the prion protein (PrP C ) from a largely a-helical isoform to a b-sheet rich oligomer (PrP Sc ). Flexibility of the polypeptide could contribute to the ability of PrP C to undergo the conformational rearrangement during PrP C -PrP Sc interactions, which then leads to the misfolded isoform. We have therefore examined the molecular motions of mouse PrP C , residues 113-231, in solution, using 15 N NMR relaxation measurements. A truncated fragment has been used to eliminate the effect of the 90-residue unstructured tail of PrP C so the dynamics of the structured domain can be studied in isolation. 15 N longitudinal (T 1 ) and transverse relaxation (T 2 ) times as well as the proton-nitrogen nuclear Overhauser effects have been used to calculate the spectral density at three frequencies, 0, x N, and 0.87x H . Spectral densities at each residue indicate various time-scale motions of the main-chain. Even within the structured domain of PrP C , a diverse range of motions are observed. We find that removal of the tail increases T 2 relaxation times significantly indicating that the tail is responsible for shortening of T 2 times in full-length PrP C . The truncated fragment of PrP has facilitated the determination of meaningful order parameters (S 2 ) from the relaxation data and shows for the first time that all three helices in PrP C have similar rigidity. Slow conformational fluctuations of mouse PrP C are localized to a distinct region that involves residues 171 and 172. Interestingly, residues 170-175 have been identified as a segment within PrP that will form a steric zipper, believed to be the fundamental amyloid unit. The flexibility within these residues could facilitate the PrP C -PrP Sc recognition process during fibril elongation.

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Section: Prp C Folding Intermediates and Stabilitymentioning

confidence: 99%

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

et al. 2009

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“…However, insights into the normal function of PrP C , and the extent to which conformational alterations might participate in this function, have proven hard to come by because of difficulties in connecting in vitro and in vivo data. For example, in vitro protein binding partners have proven difficult to authenticate by in vivo genetic analyses, and conversely, in vivo genetic analyses of PrP C have been limited to defining determinants required for prion replication and binding to an as yet uncloned partner protein designated protein X (2,3). Other recent areas of interest and controversy concern the basis of neurotoxicity and the existence and function of cytoplasmic forms of PrP C (4 -9).…”

mentioning

confidence: 99%

Genetic Mapping of Activity Determinants within Cellular Prion Proteins

Drisaldi

Coomaraswamy

Mastrangelo

et al. 2004

Journal of Biological Chemistry

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The PrP-like Doppel (Dpl) protein causes apoptotic death of cerebellar neurons in transgenic mice, a process prevented by expression of the wild type (wt) cellular prion protein, PrP C . Internally deleted forms of PrP C resembling Dpl such as PrP⌬32-121 produce a similar PrP C -sensitive pro-apoptotic phenotype in transgenic mice. Here we demonstrate that these phenotypic attributes of wt Dpl, wt PrP C , and PrP⌬132-121 can be accurately recapitulated by transfected mouse cerebellar granule cell cultures. This system was then explored by mutagenesis of the co-expressed prion proteins to reveal functional determinants. By this means, neuroprotective activity of wt PrP C was shown to be nullified by a deletion of the N-terminal charged region implicated in endocytosis and retrograde axonal transport (PrP⌬23-28), by deletion of all five octarepeats (PrP⌬51-90), or by glycine replacement of four octarepeat histidine residues required for selective binding of copper ions (Prnp"H/G"). In the case of Dpl, overlapping deletions defined a requirement for the gene interval encoding helices B and B (Dpl⌬101-125). These data suggest contributions of copper binding and neuronal trafficking to wt PrP C function in vivo and place constraints upon current hypotheses to explain Dpl/PrP C antagonism by competitive ligand binding. Further implementation of this assay should provide a fuller understanding of the attributes and subcellular localizations required for activity of these enigmatic proteins.

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“…Investigations have also revealed that the S2-H2 loop plays a principal role in the pattern-driven propagation of a prion (Kaneko et al 1997), acts as a motif with selfaggregation properties and is a universal interference target for familial prion diseases, but mutations cause the loss of the binding sites for inhibitors and the gain of an interacting region at the amyloid-forming S2-H2 loop (Meli et al 2011). Alteration of conformation of the S2-H2 loop region leads to the exposure of hydrophobic cluster to solvent, which facilitates intermolecular interactions involved in spontaneous generation of PrPSc (Biljan et al 2011).…”

Section: Resultsmentioning

confidence: 99%

Mutation in a valine residue induces drastic changes in 3D structure of human prion protein

Behmard

Abdolmaleki

Barzegari

2012

Frontiers in Life Science

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Misfolding and aggregation of the prion protein (PrP) result in prion diseases, which can occur infectiously, genetically and sporadically in humans. V210I is one of many disease-associated missense mutants of PrP, which occurs within the hydrophobic core of the protein. In the present study, we have performed molecular dynamic simulations of this PrP mutant in order to compare its dynamics and structural conformations with those of the wild-type PrP. Performed simulations illuminate the changes that occur when valine is substituted by isoleucine at position 210 of PrP. Changes can occur in helix 1 (H1), helix 2 (H2), helix 3 (H3) and around the mutation site. The V 210I mutation, especially, leads to increased flexibility in parts of the H1, H2, H3 structures that are normally stable. Overall, V 210I mutation in the hydrophobic core significantly affects the dynamics and stability of PrP. The simultaneous changes in different secondary structure elements together with perturbations in the hydrophobic core induce the drastic changes in the overall 3D structure of human prion protein.

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Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation

Cited by 469 publications

References 44 publications

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Genetic Mapping of Activity Determinants within Cellular Prion Proteins

Mutation in a valine residue induces drastic changes in 3D structure of human prion protein

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Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation

Cited by 469 publications

References 44 publications

Dynamics of a truncated prion protein, PrP(113–231), from 15N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Dynamics of a truncated prion protein, PrP(113–231), from 15N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Genetic Mapping of Activity Determinants within Cellular Prion Proteins

Mutation in a valine residue induces drastic changes in 3D structure of human prion protein

Contact Info

Product

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About

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Dynamics of a truncated prion protein, PrP(113–231), from ¹⁵N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region