title: Na+/K+-ATPase is present in scrapie-associated fibrils, modulates PrP misfolding in vitro and links PrP function and dysfunction.
abstract: Transmissible spongiform encephalopathies are characterised by widespread deposition of fibrillar and/or plaque-like forms of the prion protein. These aggregated forms are produced by misfolding of the normal prion protein, PrP(C), to the disease-associated form, PrP(Sc), through mechanisms that remain elusive but which require either direct or indirect interaction between PrP(C) and PrP(Sc) isoforms. A wealth of evidence implicates other non-PrP molecules as active participants in the misfolding process, to catalyse and direct the conformational conversion of PrP(C) or to provide a scaffold ensuring correct alignment of PrP(C) and PrP(Sc) during conversion. Such molecules may be specific to different scrapie strains to facilitate differential prion protein misfolding. Since molecular cofactors may become integrated into the growing protein fibril during prion conversion, we have investigated the proteins contained in prion disease-specific deposits by shotgun proteomics of scrapie-associated fibrils (SAF) from mice infected with 3 different strains of mouse-passaged scrapie. Concomitant use of negative control preparations allowed us to identify and discount proteins that are enriched non-specifically by the SAF isolation protocol. We found several proteins that co-purified specifically with SAF from infected brains but none of these were reproducibly and demonstrably specific for particular scrapie strains. The α-chain of Na(+)/K(+)-ATPase was common to SAF from all 3 strains and we tested the ability of this protein to modulate in vitro misfolding of recombinant PrP. Na(+)/K(+)-ATPase enhanced the efficiency of disease-specific conversion of recombinant PrP suggesting that it may act as a molecular cofactor. Consistent with previous results, the same protein inhibited fibrillisation kinetics of recombinant PrP. Since functional interactions between PrP(C) and Na(+)/K(+)-ATPase have previously been reported in astrocytes, our data highlight this molecule as a key link between PrP function, dysfunction and misfolding.
authors.affiliation: The Roslin Institute and R(D)SVS, Neuropathogenesis Division, University of Edinburgh, Easter Bush, Roslin, Edinburgh, Midlothian, United Kingdom.
authors.forename: James F
authors.initials: JF
authors.lastname: Graham
authors.forename: Dominic
authors.initials: D
authors.lastname: Kurian
authors.forename: Sonya
authors.initials: S
authors.lastname: Agarwal
authors.forename: Lorna
authors.initials: L
authors.lastname: Toovey
authors.forename: Lawrence
authors.initials: L
authors.lastname: Hunt
authors.forename: Louise
authors.initials: L
authors.lastname: Kirby
authors.forename: Teresa J T
authors.initials: TJ
authors.lastname: Pinheiro
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Banner
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Gill
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D018620:PrPSc Proteins / Q000378:metabolism*; D017510:Protein Folding; D015203:Reproducibility of Results; D012608:Scrapie / Q000201:enzymology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013045:Species Specificity; D053719:Tandem Mass Spectrometry
chemical_list: D018620:PrPSc Proteins; D000254:Sodium-Potassium-Exchanging ATPase
references.citation: Methods Mol Biol. 2008;459:131-43
references.pmid: 18576153
references.citation: J Chromatogr B Biomed Appl. 1996 May 17;680(1-2):91-6
references.pmid: 8798885
references.citation: Int J Neurosci. 2003 Jul;113(7):971-87
references.pmid: 12881189
references.citation: FEBS Lett. 2010 Jun 3;584(11):2409-14
references.pmid: 20412808
references.citation: Immun Ageing. 2010 Dec 16;7 Suppl 1:S5
references.pmid: 21172064
references.citation: Nature. 1997 Apr 10;386(6625):564
references.pmid: 9121579
references.citation: Exp Cell Res. 2006 Jul 1;312(11):1950-60
references.pmid: 16603154
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 5;104(23):9741-6
references.pmid: 17535913
references.citation: J Neurosci. 2007 Nov 7;27(45):12331-40
references.pmid: 17989297
references.citation: Eur J Biochem. 2002 May;269(10):2434-9
references.pmid: 12027880
references.citation: Biochem J. 1999 Nov 15;344 Pt 1:1-5
references.pmid: 10548526
references.citation: Biol Chem. 2005 Nov;386(11):1149-55
references.pmid: 16307480
references.citation: J Gen Virol. 2005 Jan;86(Pt 1):241-246
references.pmid: 15604453
references.citation: PLoS Pathog. 2009 Nov;5(11):e1000666
references.pmid: 19936054
references.citation: Prion. 2010 Oct-Dec;4(4):235-42
references.pmid: 20864807
references.citation: J Cell Sci. 2010 Jan 15;123(Pt 2):246-55
references.pmid: 20048341
references.citation: Proteomics. 2006 Dec;6(24):6476-84
references.pmid: 17111435
references.citation: J Biol Chem. 2010 Mar 26;285(13):9868-9880
references.pmid: 20106973
references.citation: Biochem Biophys Res Commun. 1996 Feb 6;219(1):238-42
references.pmid: 8619814
references.citation: Biochim Biophys Acta. 2009 Oct;1788(10):2245-51
references.pmid: 19703409
references.citation: Nature. 1996 Apr 18;380(6575):639-42
references.pmid: 8602267
references.citation: J Neurosci. 2007 Jan 17;27(3):616-26
references.pmid: 17234593
references.citation: Neurobiol Dis. 2005 Jun-Jul;19(1-2):255-65
references.pmid: 15837581
references.citation: Folia Neuropathol. 2009;47(2):104-13
references.pmid: 19618334
references.citation: Science. 2010 Feb 26;327(5969):1132-5
references.pmid: 20110469
references.citation: Nat Neurosci. 2010 Mar;13(3):310-8
references.pmid: 20098419
references.citation: J Cell Sci. 2008 Mar 15;121(Pt 6):773-83
references.pmid: 18285446
references.citation: Epilepsia. 1999 Dec;40(12):1679-82
references.pmid: 10612329
references.citation: Eur J Neurosci. 2008 Feb;27(3):612-20
references.pmid: 18279314
references.citation: J Mol Biol. 2001 Dec 14;314(5):1209-25
references.pmid: 11743735
references.citation: EMBO J. 2011 May 18;30(10):2057-70
references.pmid: 21441896
references.citation: PLoS One. 2009;4(2):e4446
references.pmid: 19209230
references.citation: Mol Cell Neurosci. 2009 Jun;41(2):135-47
references.pmid: 19233277
references.citation: Brain Pathol. 2007 Apr;17(2):139-45
references.pmid: 17388944
references.citation: Proteomics. 2010 Aug;10(15):2858-69
references.pmid: 20518029
references.citation: Neuron. 2009 Jul 16;63(1):12-26
references.pmid: 19607789
references.citation: Biochemistry. 2010 Feb 23;49(7):1448-59
references.pmid: 20085368
references.citation: PLoS Pathog. 2009 Oct;5(10):e1000608
references.pmid: 19798432
references.citation: Mol Neurobiol. 1994 Apr-Jun;8(2-3):121-7
references.pmid: 7999308
references.citation: J Neurosci. 2005 Dec 7;25(49):11330-9
references.pmid: 16339028
references.citation: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13363-83
references.pmid: 9811807
references.citation: Brain Res Brain Res Rev. 1991 Sep-Dec;16(3):283-300
references.pmid: 1665097
references.citation: Nat Med. 1997 Dec;3(12):1383-8
references.pmid: 9396609
references.citation: Prion. 2010 Apr-Jun;4(2):94-104
references.pmid: 20647769
references.citation: J Neurosci. 2003 Feb 15;23(4):1276-86
references.pmid: 12598616
references.citation: Brain Res Mol Brain Res. 2004 Apr 29;124(1):40-50
references.pmid: 15093684
references.citation: Nat Commun. 2011;2:281
references.pmid: 21505437
references.citation: J Neurochem. 2006 Sep;98(6):1876-85
references.pmid: 16945105
references.citation: Nat Med. 1998 Oct;4(10):1157-65
references.pmid: 9771749
references.citation: Biochem J. 1999 Sep 15;342 Pt 3:605-13
references.pmid: 10477271
references.citation: J Cell Biol. 1995 Apr;129(1):121-32
references.pmid: 7698979
references.citation: Science. 2000 Sep 15;289(5486):1925-8
references.pmid: 10988071
references.citation: Nature. 1997 Dec 18-25;390(6661):684-7
references.pmid: 9414160
references.citation: Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1807-12
references.pmid: 11842207
references.citation: Cell Mol Neurobiol. 2010 Jul;30(5):653-66
references.pmid: 20041289
references.citation: J Biol Chem. 2009 Sep 4;284(36):24233-41
references.pmid: 19596861
references.citation: Prion. 2009 Oct-Dec;3(4):240-9
references.pmid: 19901559
references.citation: J Gen Virol. 2006 Dec;87(Pt 12):3747-3751
references.pmid: 17098994
references.citation: Methods Mol Biol. 2008;459:105-15
references.pmid: 18576151
references.citation: J Gen Virol. 2003 Apr;84(Pt 4):1013-1020
references.pmid: 12655105
references.citation: Protein Sci. 2004 Nov;13(11):3017-27
references.pmid: 15459333
references.citation: Physiology (Bethesda). 2008 Aug;23:205-11
references.pmid: 18697994
references.citation: Eur J Biochem. 1988 Mar 1;172(2):271-7
references.pmid: 2894984
references.citation: Amino Acids. 2008 Feb;34(2):175-85
references.pmid: 17177074
references.citation: Free Radic Biol Med. 2007 Sep 15;43(6):959-67
references.pmid: 17697940
references.citation: Hum Mol Genet. 2007 Mar 1;16(5):555-66
references.pmid: 17309878
references.citation: Proteomics. 2009 Aug;9(15):3802-14
references.pmid: 19637240
references.citation: Biochim Biophys Acta. 2008 Jun;1784(6):873-81
references.pmid: 18374666
references.citation: Med Hypotheses. 1995 Mar;44(3):173-8
references.pmid: 7609671
references.citation: J Biol Chem. 1998 Nov 27;273(48):32230-5
references.pmid: 9822701
references.citation: Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2212-7
references.pmid: 19164762
references.citation: Nature. 2003 Oct 16;425(6959):717-20
references.pmid: 14562104
references.citation: J Mol Neurosci. 2006;29(3):195-214
references.pmid: 17085779
references.citation: Prog Neurobiol. 2001 Sep;65(1):1-105
references.pmid: 11369436
references.citation: J Neurochem. 2011 Mar;116(5):881-90
references.pmid: 21214552
references.citation: J Neurochem. 2010 Jul;114(2):408-18
references.pmid: 20412382
references.citation: J Proteomics. 2009 May 2;72(4):690-4
references.pmid: 19367687