Oxidation of Helix-3 Methionines Precedes the Formation of PK Resistant PrP

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While elucidating the peculiar epitope of the α-PrP mAb IPC2, we found that PrP^Sc exhibits the sulfoxidation of residue M213 as a covalent signature. Subsequent computational analysis predicted that the presence of sulfoxide groups at both Met residues 206 and 213 destabilize the α-fold, suggesting oxidation may facilitate the conversion of PrP^C into PrP^Sc. To further study the effect of oxidation on prion formation, we generated pAbs to linear PrP peptides encompassing the Helix-3 region, as opposed to the non-linear complexed epitope of IPC2. We now show that pAbs, whose epitopes comprise Met residues, readily detected PrP^C, but could not recognize most PrP^Sc bands unless they were vigorously reduced. Next, we showed that the α-Met pAbs did not recognize newly formed PrP^Sc, as is the case for the PK resistant PrP present in lines of prion infected cells. In addition, these reagents did not detect intermediate forms such as PK sensitive and partially aggregated PrPs present in infected brains. Finally, we show that PrP molecules harboring the pathogenic mutation E200K, which is linked to the most common form of familial CJD, may be spontaneously oxidized. We conclude that the oxidation of methionine residues in Helix-3 represents an early and important event in the conversion of PrP^C to PrP^Sc. We believe that further investigation into the mechanism and role of PrP oxidation will be central in finally elucidating the mechanism by which a normal cell protein converts into a pathogenic entity that causes fatal brain degeneration.

Vyšlo v časopise: Oxidation of Helix-3 Methionines Precedes the Formation of PK Resistant PrP. PLoS Pathog 6(7): e32767. doi:10.1371/journal.ppat.1000977
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000977

Souhrn

Zdroje

1. PrusinerSB

ScottMR

DeArmondSJ

CohenFE

1998 Prion protein biology. Cell 93(3) 337 348

2. BrownDR

2005 Neurodegeneration and oxidative stress: prion disease results from loss of antioxidant defence. Folia Neuropathol 43 229 243

3. NadalRC

AbdelraheimSR

BrazierMW

RigbySE

BrownDR

2007 Prion protein does not redox-silence Cu2+, but is a sacrificial quencher of hydroxyl radicals. Free Radic Biol Med 42 79 89

4. BrownDR

CliveC

HaswellSJ

2001 Antioxidant activity related to copper binding of native prion protein. J Neurochem 76 69 76

5. BrownDR

SassoonJ

2002 Copper-dependent functions for the prion protein. Mol Biotechnol 22 165 178

6. PanKM

BaldwinM

NguyenJ

GassetM

SerbanA

1993 Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci U S A 90(23) 10962 10966

7. CanelloT

EngelsteinR

MoshelO

XanthopoulosK

JuanesME

2008 Methionine sulfoxides on PrPSc: a prion-specific covalent signature. Biochemistry 47 8866 8873

8. OienDB

CanelloT

GabizonR

GassetM

LundquistBL

2009 Detection of oxidized methionine in selected proteins, cellular extracts and blood serums by novel anti-methionine sulfoxide antibodies. Arch Biochem Biophys 485 35 40

9. StahlN

BaldwinMA

TeplowDB

HoodL

GibsonBW

1993 Structural studies of the scrapie prion protein using mass spectrometry and amino acid sequencing. Biochemistry 32 1991 2002

10. RequenaJR

GrothD

LegnameG

StadtmanER

PrusinerSB

2001 Copper-catalyzed oxidation of the recombinant SHa(29-231) prion protein. Proc Natl Acad Sci U S A 98 7170 7175

11. HirschbergerT

StorkM

SchroppB

WinklhoferKF

TatzeltJ

2006 Structural instability of the prion protein upon M205S/R mutations revealed by molecular dynamics simulations. Biophys J 90 3908 3918

12. MoskovitzJ

Bar-NoyS

WilliamsWM

RequenaJ

BerlettBS

2001 Methionine sulfoxide reductase (MsrA) is a regulator of antioxidant defense and lifespan in mammals. Proc Natl Acad Sci U S A 98 12920 12925

13. BingerKJ

GriffinMD

HeinemannSH

HowlettGJ

Methionine-oxidized amyloid fibrils are poor substrates for human methionine sulfoxide reductases A and B2. Biochemistry 49 2981 2983

14. TamguneyG

GilesK

GliddenDV

LessardP

WilleH

2008 Genes contributing to prion pathogenesis. J Gen Virol 89 1777 1788

15. ColomboG

MeliM

MorraG

GabizonR

GassetM

2009 Methionine sulfoxides on prion protein Helix-3 switch on the alpha-fold destabilization required for conversion. PLoS ONE 4 e4296

16. WolschnerC

GieseA

KretzschmarHA

HuberR

MoroderL

2009 Design of anti- and pro-aggregation variants to assess the effects of methionine oxidation in human prion protein. Proc Natl Acad Sci U S A 106 7756 7761

17. LisaS

MeliM

CabelloG

GabizonR

ColomboG

2010 The structural intolerance of the.PrP a-fold for polar substitution of the helix-3 methionines. Cell Mol Life Sci in press

18. HsiaoK

MeinerZ

KahanaE

CassC

KahanaI

1991 Mutation of the prion protein in Libyan Jews with Creutzfeldt-Jakob disease. N Engl J Med 324 1091 1097

19. WeiskopfD

SchwanningerA

WeinbergerB

AlmanzarG

ParsonW

2010 Oxidative stress can alter the antigenicity of immunodominant peptides. J Leukoc Biol 87 165 172

20. SulzerB

PerelsonAS

1997 Immunons revisited: binding of multivalent antigens to B cells. Mol Immunol 34 63 74

21. WilliamsonRA

PeretzD

PinillaC

BallH

BastidasRB

1998 Mapping the prion protein using recombinant antibodies. J Virol 72 9413 9418

22. HoughtenRA

LiCH

1979 Reduction of sulfoxides in peptides and proteins. Anal Biochem 98 36 46

23. ButlerDA

ScottMR

BockmanJM

BorcheltDR

TaraboulosA

1988 Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins. J Virol 62 1558 1564

24. LeuchtC

SimoneauS

ReyC

VanaK

RiegerR

2003 The 37 kDa/67 kDa laminin receptor is required for PrP(Sc) propagation in scrapie-infected neuronal cells. EMBO Rep 4 290 295

25. LiuY

SunR

ChakrabartyT

ManuelidisL

2008 A rapid accurate culture assay for infectivity in Transmissible Encephalopathies. J Neurovirol 14 352 361

26. ScottMR

KohlerR

FosterD

PrusinerSB

1992 Chimeric prion protein expression in cultured cells and transgenic mice. Protein Sci 1 986 997

27. CaspiS

HalimiM

YanaiA

SassonSB

TaraboulosA

1998 The anti-prion activity of Congo red. Putative mechanism. J Biol Chem 273 3484 3489

28. BorcheltDR

ScottM

TaraboulosA

StahlN

PrusinerSB

1990 Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. J Cell Biol 110 743 752

29. CaugheyB

RaceRE

ErnstD

BuchmeierMJ

ChesebroB

1989 Prion protein biosynthesis in scrapie-infected and uninfected neuroblastoma cells. J Virol 63 175 181

30. TzabanS

FriedlanderG

SchonbergerO

HoronchikL

YedidiaY

2002 Protease-sensitive scrapie prion protein in aggregates of heterogeneous sizes. Biochemistry 41 12868 12875

31. SafarJ

CeroniM

PiccardoP

LiberskiPP

MiyazakiM

1990 Subcellular distribution and physicochemical properties of scrapie-associated precursor protein and relationship with scrapie agent. Neurology 40 503 508

32. SafarJ

WilleH

ItriV

GrothD

SerbanH

1998 Eight prion strains have PrP(Sc) molecules with different conformations. Nat Med 4 1157 1165

33. SerbanD

TaraboulosA

DeArmondSJ

PrusinerSB

1990 Rapid detection of Creutzfeldt-Jakob disease and scrapie prion proteins. Neurology 40 110 117

34. GabizonR

TellingG

MeinerZ

HalimiM

KahanaI

1996 Insoluble wild-type and protease-resistant mutant prion protein in brains of patients with inherited prion disease. Nat Med 2 59 64

35. ChenSG

ParchiP

BrownP

CapellariS

ZouW

1997 Allelic origin of the abnormal prion protein isoform in familial prion diseases. Nat Med 3 1009 1015

36. SwietnickiW

PetersenRB

GambettiP

SurewiczWK

1998 Familial mutations and the thermodynamic stability of the recombinant human prion protein. J Biol Chem 273 31048 31052

37. ZhangY

SwietnickiW

ZagorskiMG

SurewiczWK

SonnichsenFD

2000 Solution structure of the E200K variant of human prion protein. Implications for the mechanism of pathogenesis in familial prion diseases. J Biol Chem 275 33650 33654

38. WinklhoferKF

HeskeJ

HellerU

ReintjesA

MuranyiW

2003 Determinants of the in vivo folding of the prion protein. A bipartite function of helix 1 in folding and aggregation. J Biol Chem 278 14961 14970

39. HartT

HosszuLL

TrevittCR

JacksonGS

WalthoJP

2009 Folding kinetics of the human prion protein probed by temperature jump. Proc Natl Acad Sci U S A 106 5651 5656

40. SilvaCJ

OniskoBC

DyninI

EricksonML

VenselWH

Assessing the role of oxidized methionine at position 213 in the formation of prions in hamsters. Biochemistry 49 1854 1861

41. FroelichJM

ReidGE

2008 The origin and control of ex vivo oxidative peptide modifications prior to mass spectrometry analysis. Proteomics 8 1334 1345

42. OniskoB

DyninI

RequenaJR

SilvaCJ

EricksonM

2007 Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS. J Am Soc Mass Spectrom 18 1070 1079

43. SpudichS

MastrianniJA

WrenschM

GabizonR

MeinerZ

1995 Complete penetrance of Creutzfeldt-Jakob disease in Libyan Jews carrying the E200K mutation in the prion protein gene. Mol Med 1 607 613

44. ChapmanJ

Ben-IsraelJ

GoldhammerY

KorczynAD

1994 The risk of developing Creutzfeldt-Jakob disease in subjects with the PRNP gene codon 200 point mutation. Neurology 44 1683 1686

45. StadtmanER

BerlettBS

1997 Reactive oxygen-mediated protein oxidation in aging and disease. Chem Res Toxicol 10 485 494

46. MakaravaN

KovacsGG

BocharovaO

SavtchenkoR

AlexeevaI

2010 Recombinant prion protein induces a new transmissible prion disease in wild-type animals. Acta Neuropathol 119 177 187

47. LegnameG

BaskakovIV

NguyenHO

RiesnerD

CohenFE

2004 Synthetic mammalian prions. Science 305 673 676

48. SaborioGP

PermanneB

SotoC

2001 Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature 411 810 813

49. AtarashiR

MooreRA

SimVL

HughsonAG

DorwardDW

2007 Ultrasensitive detection of scrapie prion protein using seeded conversion of recombinant prion protein. Nat Methods 4 645 650

50. WangF

WangX

YuanCG

MaJ

Generating a prion with bacterially expressed recombinant prion protein. Science 327 1132 1135

51. BuelerH

FischerM

LangY

BluethmannH

LippHP

1992 Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein [see comments]. Nature 356 577 582

52. VaradarajanS

YatinS

AksenovaM

ButterfieldDA

2000 Review: Alzheimer's amyloid beta-peptide-associated free radical oxidative stress and neurotoxicity. J Struct Biol 130 184 208

53. ChiL

KeY

LuoC

GozalD

LiuR

2007 Depletion of reduced glutathione enhances motor neuron degeneration in vitro and in vivo. Neuroscience 144 991 1003

54. Gonzalez-IglesiasR

PajaresMA

OcalC

EspinosaJC

OeschB

2002 Prion protein interaction with glycosaminoglycan occurs with the formation of oligomeric complexes stabilized by Cu(II) bridges. J Mol Biol 319 527 540

55. MakaravaN

BaskakovIV

2008 Expression and purification of full-length recombinant PrP of high purity. Methods Mol Biol 459 131 143