title: Glycosaminoglycan sulphation affects the seeded misfolding of a mutant prion protein.
abstract: BACKGROUND
  The accumulation of protease resistant conformers of the prion protein (PrP(res)) is a key pathological feature of prion diseases. Polyanions, including RNA and glycosaminoglycans have been identified as factors that contribute to the propagation, transmission and pathogenesis of prion disease. Recent studies have suggested that the contribution of these cofactors to prion propagation may be species specific.
  
  
  RESULTS
  In this study a cell-free assay was used to investigate the molecular basis of polyanion stimulated PrP(res) formation using brain tissue or cell line derived murine PrP. Enzymatic depletion of endogenous nucleic acids or heparan sulphate (HS) from the PrP(C) substrate was found to specifically prevent PrP(res) formation seeded by mouse derived PrP(Sc). Modification of the negative charge afforded by the sulphation of glycosaminoglycans increased the ability of a familial PrP mutant to act as a substrate for PrP(res) formation, while having no effect on PrP(res) formed by wildtype PrP. This difference may be due to the observed differences in the binding of wild type and mutant PrP for glycosaminoglycans.
  
  
  CONCLUSIONS
  Cofactor requirements for PrP(res) formation are host species and prion strain specific and affected by disease associated mutations of the prion protein. This may explain both species and strain dependent propagation characteristics and provide insights into the underlying mechanisms of familial prion disease. It further highlights the challenge of designing effective therapeutics against a disease which effects a range of mammalian species, caused by range of aetiologies and prion strains.
authors.affiliation: Department of Pathology, The University of Melbourne, Parkville, Victoria, Australia. vlawson@unimelb.edu.au
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Lawson
authors.forename: Brooke
authors.initials: B
authors.lastname: Lumicisi
authors.forename: Jeremy
authors.initials: J
authors.lastname: Welton
authors.forename: Dorothy
authors.initials: D
authors.lastname: Machalek
authors.forename: Katrina
authors.initials: K
authors.lastname: Gouramanis
authors.forename: Helen M
authors.initials: HM
authors.lastname: Klemm
authors.forename: James D
authors.initials: JD
authors.lastname: Stewart
authors.forename: Colin L
authors.initials: CL
authors.lastname: Masters
authors.forename: David E
authors.initials: DE
authors.lastname: Hoke
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Collins
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Hill
mesh_terms: D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D051379:Mice; D050505:Mutant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009696:Nucleic Acids / Q000378:metabolism; D017096:Prion Diseases / Q000235:genetics / Q000378:metabolism; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D055672:Static Electricity; D013431:Sulfates / Q000378:metabolism*
chemical_list: D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D050505:Mutant Proteins; D009696:Nucleic Acids; D011328:Prions; D013431:Sulfates
references.citation: Brain Res Bull. 2006 Jan 30;68(5):346-54
references.pmid: 16377442
references.citation: Cell. 2005 Apr 22;121(2):195-206
references.pmid: 15851027
references.citation: Ann Neurol. 1979 Jun;5(6):581-4
references.pmid: 382976
references.citation: J Mol Biol. 2004 Nov 26;344(3):827-37
references.pmid: 15533448
references.citation: J Pathol. 2007 Apr;211(5):582-590
references.pmid: 17334982
references.citation: J Biol Chem. 2005 Apr 29;280(17):17062-7
references.pmid: 15668247
references.citation: Biochem Biophys Res Commun. 1986 Dec 15;141(2):870-7
references.pmid: 3026396
references.citation: Science. 2004 Jul 30;305(5684):673-6
references.pmid: 15286374
references.citation: J Recept Signal Transduct Res. 2008;28(3):153-61
references.pmid: 18569523
references.citation: J Mol Biol. 2001 Apr 6;307(4):1011-21
references.pmid: 11286552
references.citation: J Mol Biol. 2010 Jan 22;395(3):475-90
references.pmid: 19913031
references.citation: Arch Virol Suppl. 2000;(16):95-102
references.pmid: 11214938
references.citation: Biochem Biophys Res Commun. 2004 Jan 16;313(3):559-67
references.pmid: 14697227
references.citation: Nature. 1994 Aug 11;370(6489):471-4
references.pmid: 7913989
references.citation: Lancet. 2004 Jan 3;363(9402):51-61
references.pmid: 14723996
references.citation: Biochem Soc Symp. 2005;(72):211-22
references.pmid: 15649144
references.citation: C R Biol. 2002 Jan;325(1):17-23
references.pmid: 11862616
references.citation: Science. 2010 Feb 26;327(5969):1132-5
references.pmid: 20110469
references.citation: J Gen Virol. 2008 Jan;89(Pt 1):341-347
references.pmid: 18089759
references.citation: EMBO J. 2001 Feb 1;20(3):377-86
references.pmid: 11157745
references.citation: Lab Invest. 1990 Nov;63(5):601-11
references.pmid: 1977959
references.citation: Biochemistry. 2001 Jun 12;40(23):6982-7
references.pmid: 11389614
references.citation: Biochemistry. 2003 Mar 25;42(11):3295-304
references.pmid: 12641461
references.citation: J Mol Biol. 2002 Sep 6;322(1):153-61
references.pmid: 12215421
references.citation: J Biol Chem. 2004 Sep 24;279(39):40788-94
references.pmid: 15262998
references.citation: Nature. 2001 Jun 14;411(6839):810-3
references.pmid: 11459061
references.citation: Nature. 2006 Oct 19;443(7113):803-10
references.pmid: 17051207
references.citation: Proc Natl Acad Sci U S A. 2007 May 1;104(18):7546-51
references.pmid: 17456603
references.citation: Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4055-9
references.pmid: 11259656
references.citation: Biochemistry. 2010 May 11;49(18):3928-34
references.pmid: 20377181
references.citation: J Mol Biol. 2003 Sep 5;332(1):47-57
references.pmid: 12946346
references.citation: J Pathol Bacteriol. 1968 Apr;95(2):377-81
references.pmid: 4297026
references.citation: Arch Biochem Biophys. 1999 Oct 15;370(2):250-7
references.pmid: 10510284
references.citation: J Recept Signal Transduct Res. 2007;27(5-6):433-43
references.pmid: 18097941
references.citation: Protein Sci. 2006 Mar;15(3):609-19
references.pmid: 16452616
references.citation: J Virol. 2004 May;78(10):4999-5006
references.pmid: 15113880
references.citation: J Biol Chem. 1995 Dec 15;270(50):30221-9
references.pmid: 8530433
references.citation: Acta Neuropathol. 1989;77(4):337-42
references.pmid: 2523631
references.citation: J Gen Virol. 2003 Sep;84(Pt 9):2595-2603
references.pmid: 12917481
references.citation: J Biol Chem. 2002 May 24;277(21):18421-30
references.pmid: 11882649
references.citation: J Mol Biol. 2002 May 31;319(2):527-40
references.pmid: 12051926
references.citation: J Biol Chem. 2005 Jul 22;280(29):26873-9
references.pmid: 15917229
references.citation: Biochemistry. 2003 Apr 15;42(14):4127-35
references.pmid: 12680767
references.citation: J Neurol Neurosurg Psychiatry. 2007 Jul;78(7):733-4
references.pmid: 17314188
references.citation: J Biol Chem. 1996 Jan 19;271(3):1633-7
references.pmid: 8576163
references.citation: J Cell Physiol. 1993 Nov;157(2):319-25
references.pmid: 7901226
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 5;104(23):9551-2
references.pmid: 17535888
references.citation: J Biol Chem. 2005 Apr 29;280(17):17057-61
references.pmid: 15668233
references.citation: J Gen Virol. 2007 Oct;88(Pt 10):2905-2914
references.pmid: 17872546
references.citation: J Mol Biol. 2002 Feb 1;315(5):1241-56
references.pmid: 11827491
references.citation: Chembiochem. 2002 Aug 2;3(8):717-25
references.pmid: 12203970
references.citation: EMBO J. 1996 Mar 15;15(6):1255-64
references.pmid: 8635458
references.citation: J Biol Chem. 2003 Oct 10;278(41):40041-9
references.pmid: 12871949
references.citation: J Biol Chem. 2010 May 7;285(19):14083-7
references.pmid: 20304915
references.citation: Cell. 1993 Jul 2;73(7):1339-47
references.pmid: 8100741
references.citation: J Virol. 2007 Oct;81(19):10786-91
references.pmid: 17626095
references.citation: Biochemistry. 1999 Mar 16;38(11):3280-4
references.pmid: 10079070
references.citation: Eur J Neurol. 2008 May;15(5):458-64
references.pmid: 18355301
references.citation: Science. 1982 Apr 9;216(4542):136-44
references.pmid: 6801762
references.citation: J Biol Chem. 1999 Dec 17;274(51):36267-73
references.pmid: 10593915
references.citation: Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11667-72
references.pmid: 9751723
references.citation: J Recept Signal Transduct Res. 2007;27(5-6):411-31
references.pmid: 18097940
references.citation: Nature. 2003 Oct 16;425(6959):717-20
references.pmid: 14562104
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 5;104(23):9741-6
references.pmid: 17535913
references.citation: Int J Biochem Cell Biol. 2008;40(12):2793-801
references.pmid: 18590830
references.citation: J Biol Chem. 1998 Nov 20;273(47):31048-52
references.pmid: 9813003
references.citation: J Biol Chem. 2001 Jun 1;276(22):19301-9
references.pmid: 11278562