skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Differential effects of divalent cations on elk prion protein fibril formation and stability

Abstract

Misfolding of the normally folded prion protein of mammals (PrP C) into infectious fibrils causes a variety of diseases, from scrapie in sheep to chronic wasting disease (CWD) in cervids. The misfolded form of PrP C, termed PrP Sc, or in this case PrP CWD, interacts with PrP C to create more PrP CWD. This process is not clearly defined but is affected by the presence and interactions of biotic and abiotic cofactors. These include nucleic acids, lipids, glycosylation, pH, and ionic character. PrP C has been shown to act as a copper-binding protein in vivo, though it also binds to other divalents as well. The significance of this action has not been conclusively elucidated. Previous reports have shown that metal binding sites occur throughout the N-terminal region of PrP C. Other cations like manganese have also been shown to affect PrP C abundance in a transcript-independent fashion. Here, we examined the ability of different divalent cations to influence the stability and in vitro conversion of two variants of PrP from elk (L/M132, 26–234). We find that copper and zinc destabilize PrP. We also find that PrP M132 exhibits a greater degree of divalent cation induced destabilization than L132. Thismore » supports findings that leucine at position 132 confers resistance to CWD, while M132 is susceptible. However, in vitro conversion of PrP is equally suppressed by either copper or zinc, in both L132 and M132 backgrounds. This report demonstrates the complex importance of ionic character on the PrP C folding pathway selection on the route to PrP Sc formation.« less

Authors:
 [1];  [2]
  1. Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States); United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, IA (United States)
  2. United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, IA (United States)
Publication Date:
Research Org.:
Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1494691
Grant/Contract Number:  
AC05-06OR23100
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Prion
Additional Journal Information:
Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 1933-6896
Publisher:
Taylor & Francis
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Prion; chronic wasting disease; PrP; transmissible spongiform encephalopathies; cervid; CWD; amyloid; misfolding

Citation Formats

Samorodnitsky, Daniel, and Nicholson, Eric M. Differential effects of divalent cations on elk prion protein fibril formation and stability. United States: N. p., 2018. Web. doi:10.1080/19336896.2017.1423187.
Samorodnitsky, Daniel, & Nicholson, Eric M. Differential effects of divalent cations on elk prion protein fibril formation and stability. United States. doi:10.1080/19336896.2017.1423187.
Samorodnitsky, Daniel, and Nicholson, Eric M. Tue . "Differential effects of divalent cations on elk prion protein fibril formation and stability". United States. doi:10.1080/19336896.2017.1423187. https://www.osti.gov/servlets/purl/1494691.
@article{osti_1494691,
title = {Differential effects of divalent cations on elk prion protein fibril formation and stability},
author = {Samorodnitsky, Daniel and Nicholson, Eric M.},
abstractNote = {Misfolding of the normally folded prion protein of mammals (PrPC) into infectious fibrils causes a variety of diseases, from scrapie in sheep to chronic wasting disease (CWD) in cervids. The misfolded form of PrPC, termed PrPSc, or in this case PrPCWD, interacts with PrPC to create more PrPCWD. This process is not clearly defined but is affected by the presence and interactions of biotic and abiotic cofactors. These include nucleic acids, lipids, glycosylation, pH, and ionic character. PrPC has been shown to act as a copper-binding protein in vivo, though it also binds to other divalents as well. The significance of this action has not been conclusively elucidated. Previous reports have shown that metal binding sites occur throughout the N-terminal region of PrPC. Other cations like manganese have also been shown to affect PrPC abundance in a transcript-independent fashion. Here, we examined the ability of different divalent cations to influence the stability and in vitro conversion of two variants of PrP from elk (L/M132, 26–234). We find that copper and zinc destabilize PrP. We also find that PrP M132 exhibits a greater degree of divalent cation induced destabilization than L132. This supports findings that leucine at position 132 confers resistance to CWD, while M132 is susceptible. However, in vitro conversion of PrP is equally suppressed by either copper or zinc, in both L132 and M132 backgrounds. This report demonstrates the complex importance of ionic character on the PrPC folding pathway selection on the route to PrPSc formation.},
doi = {10.1080/19336896.2017.1423187},
journal = {Prion},
issn = {1933-6896},
number = 1,
volume = 12,
place = {United States},
year = {2018},
month = {1}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 3 works
Citation information provided by
Web of Science

Figures / Tables:

Figure 1 Figure 1: Determination of Tm of Elk rPrP variants by thermal denaturation circular dichroism spectroscopy. Both rPrP L132 (A) and M132 (B) were melted in either the absence or presence of increasing molar ratios of divalent cations, copper (black circles), manganese (pink squares), calcium (blue triangles), or zinc (purple triangles).more » In the absence of divalents, both variants have similar melting temperatures – 67.16°C for L132 and 68.08°C for M132. The profile of M132 in the presence of 50x copper did not exhibit a sigmoidal shaped unfolding curve consistent with two state cooperative unfolding and could not be used to determine a Tm and is not included in this figure. In the presence of 120 mM NaCl, L132’s Tm was 64.580C and M132’s was 68.01°C.« less

Save / Share:

Works referenced in this record:

Prion protein NMR structures of elk and of mouse/elk hybrids
journal, January 2005

  • Gossert, A. D.; Bonjour, S.; Lysek, D. A.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 3
  • DOI: 10.1073/pnas.0409008102

Dietary magnesium and copper affect survival time and neuroinflammation in chronic wasting disease
journal, May 2016


Cross-species transmission of CWD prions
journal, January 2016


Experimental Oral Transmission of Chronic Wasting Disease to Reindeer (Rangifer tarandus tarandus)
journal, June 2012


First case of chronic wasting disease in Europe in a Norwegian free-ranging reindeer
journal, September 2016

  • Benestad, Sylvie L.; Mitchell, Gordon; Simmons, Marion
  • Veterinary Research, Vol. 47, Issue 1
  • DOI: 10.1186/s13567-016-0375-4

Methods to Estimate the Conformation of Proteins and Polypeptides from Circular Dichroism Data
journal, March 1996


Zinc Drives a Tertiary Fold in the Prion Protein with Familial Disease Mutation Sites at the Interface
journal, February 2013


The Priori Diseases
journal, July 1998


Spongiform Encephalopathy of Rocky Mountain elk
journal, October 1982


Thermodynamics of unfolding for turkey ovomucoid third domain: Thermal and chemical denaturation
journal, December 1993


The Prion Protein is a Combined Zinc and Copper Binding Protein:  Zn 2+ Alters the Distribution of Cu 2+ Coordination Modes
journal, December 2007

  • Walter, Eric D.; Stevens, Daniel J.; Visconte, Micah P.
  • Journal of the American Chemical Society, Vol. 129, Issue 50
  • DOI: 10.1021/ja077146j

Chronic Wasting Disease of Captive mule Deer: a Spongiform Encephalopathy
journal, January 1980



journal, October 2009

  • Walter, Eric; Stevens, Daniel; Spevacek, Ann
  • Current Protein & Peptide Science, Vol. 10, Issue 5
  • DOI: 10.2174/138920309789352056

Prion Seeding Activities of Mouse Scrapie Strains with Divergent PrPSc Protease Sensitivities and Amyloid Plaque Content Using RT-QuIC and eQuIC
journal, November 2012


Comparison of Two US Sheep Scrapie Isolates Supports Identification as Separate Strains
journal, July 2016


Dissociation of pathological and molecular phenotype of variant Creutzfeldt-Jakob disease in transgenic human prion protein 129 heterozygous mice
journal, June 2006

  • Asante, E. A.; Linehan, J. M.; Gowland, I.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 28
  • DOI: 10.1073/pnas.0604292103

Novel proteinaceous infectious particles cause scrapie
journal, April 1982


Copper(II) Inhibits in Vitro Conversion of Prion Protein into Amyloid Fibrils
journal, May 2005

  • Bocharova, Olga V.; Breydo, Leonid; Salnikov, Vadim V.
  • Biochemistry, Vol. 44, Issue 18
  • DOI: 10.1021/bi050251q

Prion Protein Interaction with Glycosaminoglycan Occurs with the Formation of Oligomeric Complexes Stabilized by Cu(II) Bridges
journal, May 2002

  • González-Iglesias, Reinerio; Pajares, Marı́a A.; Ocal, Carmen
  • Journal of Molecular Biology, Vol. 319, Issue 2
  • DOI: 10.1016/S0022-2836(02)00341-8

Resistance to Chronic Wasting Disease in Transgenic Mice Expressing a Naturally Occurring Allelic Variant of Deer Prion Protein
journal, February 2007

  • Meade-White, K.; Race, B.; Trifilo, M.
  • Journal of Virology, Vol. 81, Issue 9
  • DOI: 10.1128/JVI.02762-06

Molecular mechanism of Thioflavin-T binding to amyloid fibrils
journal, July 2010

  • Biancalana, Matthew; Koide, Shohei
  • Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, Vol. 1804, Issue 7
  • DOI: 10.1016/j.bbapap.2010.04.001

Contributions of the Prion Protein Sequence, Strain, and Environment to the Species Barrier
journal, November 2015

  • Sharma, Aditi; Bruce, Kathryn L.; Chen, Buxin
  • Journal of Biological Chemistry, Vol. 291, Issue 3
  • DOI: 10.1074/jbc.M115.684100

Metal binding sites in amyloid oligomers: Complexes and mechanisms
journal, October 2012

  • Miller, Yifat; Ma, Buyong; Nussinov, Ruth
  • Coordination Chemistry Reviews, Vol. 256, Issue 19-20
  • DOI: 10.1016/j.ccr.2011.12.022

Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays
journal, December 2010


Strain Characterization of the Korean CWD Cases in 2001 and 2004
journal, January 2013

  • Lee, Yoon-Hee; Sohn, Hyun-Joo; Kim, Min-Jeong
  • Journal of Veterinary Medical Science, Vol. 75, Issue 1
  • DOI: 10.1292/jvms.12-0077

Prion Protein Selectively Binds Copper(II) Ions
journal, May 1998

  • Stöckel, Johannes; Safar, Jiri; Wallace, Andrew C.
  • Biochemistry, Vol. 37, Issue 20
  • DOI: 10.1021/bi972827k

In vitro Conversion of Full-length Mammalian Prion Protein Produces Amyloid Form with Physical Properties of PrPSc
journal, February 2005

  • Bocharova, Olga V.; Breydo, Leonid; Parfenov, Alexander S.
  • Journal of Molecular Biology, Vol. 346, Issue 2
  • DOI: 10.1016/j.jmb.2004.11.068

A comparative analysis of rapid methods for purification and refolding of recombinant bovine prion protein
journal, April 2012

  • Vrentas, Catherine E.; Onstot, Stephanie; Nicholson, Eric M.
  • Protein Expression and Purification, Vol. 82, Issue 2
  • DOI: 10.1016/j.pep.2012.02.008

Human Prion Protein with Valine 129 Prevents Expression of Variant CJD Phenotype
journal, November 2004

  • Wadsworth, Jonathan D. F.; Asante, Emmanuel A.; Desbruslais, Melanie
  • Science, Vol. 306, Issue 5702
  • DOI: 10.1126/science.1103932

Preliminary Observations of Genetic Susceptibility of elk ( Cervus Elaphus Nelsoni ) to Chronic Wasting Disease by Experimental Oral Inoculation
journal, January 2006

  • Hamir, Amir N.; Gidlewski, Thomas; Spraker, Terry R.
  • Journal of Veterinary Diagnostic Investigation, Vol. 18, Issue 1
  • DOI: 10.1177/104063870601800118

Elk with a long incubation prion disease phenotype have a unique PrPd profile
journal, January 2007


Molecular-Level Examination of Cu 2+ Binding Structure for Amyloid Fibrils of 40-Residue Alzheimer’s β by Solid-State NMR Spectroscopy
journal, March 2011

  • Parthasarathy, Sudhakar; Long, Fei; Miller, Yifat
  • Journal of the American Chemical Society, Vol. 133, Issue 10
  • DOI: 10.1021/ja1072178

Trans-species amplification of PrPCWD and correlation with rigid loop 170N
journal, April 2009


Protein secondary structure and circular dichroism: A practical guide
journal, January 1990


Zinc ions promote Alzheimer A  aggregation via population shift of polymorphic states
journal, May 2010

  • Miller, Y.; Ma, B.; Nussinov, R.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 21
  • DOI: 10.1073/pnas.0913114107

Using circular dichroism spectra to estimate protein secondary structure
journal, January 2007


    Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.