JBIC Journal of Biological Inorganic Chemistry

, Volume 9, Issue 8, pp 954–960 | Cite as

Trace metal contamination initiates the apparent auto-aggregation, amyloidosis, and oligomerization of Alzheimer’s Aβ peptides

  • Xudong Huang
  • Craig S. Atwood
  • Robert D. Moir
  • Mariana A. Hartshorn
  • Rudolph E. Tanzi
  • Ashley I. Bush
Original Article


Nucleation-dependent protein aggregation (“seeding”) and amyloid fibril-free formation of soluble SDS-resistant oligomers (“oligomerization”) by hydrophobic interaction is an in vitro model thought to propagate β-amyloid (Aβ) deposition, accumulation, and incur neurotoxicity and synaptotoxicity in Alzheimer’s disease (AD), and other amyloid-associated neurodegenerative diseases. However, Aβ is a high-affinity metalloprotein that aggregates in the presence of biometals (zinc, copper, and iron), and neocortical Aβ deposition is abolished by genetic ablation of synaptic zinc in transgenic mice. We now present in vitro evidence that trace (≤0.8 µM) levels of zinc, copper, and iron, present as common contaminants of laboratory buffers and culture media, are the actual initiators of the classic Aβ1–42-mediated seeding process and Aβ oligomerization. Replicating the experimental conditions of earlier workers, we found that the in vitro precipitation and amyloidosis of Aβ1–40 (20 µM) initiated by Aβ1–42 (2 µM) were abolished by chelation of trace metal contaminants. Further, metal chelation attenuated formation of soluble Aβ oligomers from a cell-free culture medium. These data suggest that protein self-assembly and oligomerization are not spontaneous in this system as previously thought, and that there may be an obligatory role for metal ions in initiating Aβ amyloidosis and oligomerization.


Aβ amyloid Aβ oligomerization Alzheimer’s disease Metal chelator Trace metal contaminants 




Alzheimer’s disease


Congo Red


Dulbecco’s modified Eagle’s medium


diethylenetriaminepentaacetic acid


ion coupled plasma-mass spectroscopy


sodium dodecyl sulfate-polyacrylamide gel electrophoresis





X.H. was supported by a NRSA award (5F32AG05782) from the NIH, and is a recipient of an NIH Career Development grant (5K01MH02001) and an AFAR grant. A.I.B. was supported by funds from the NIA (5R01AG12685), the American Health Assistance Foundation, the Alzheimer Association, and the Neuroscience Research and Education Foundation. R.E.T. is an Ellison Medical Foundation Senior Scholar.


  1. 1.
    Glenner GG, Wong CW (1984) Biochem Biophys Res Commun 120:885–890PubMedGoogle Scholar
  2. 2.
    Vigo-Pelfrey C, Lee D, Keim P, Lieberburg I, Schenk DB (1993) J Neurochem 61:1965–1968PubMedGoogle Scholar
  3. 3.
    Prelli F, Casta E, Glenner GG, Frangione B (1988) J Neurochem 51:648–651PubMedGoogle Scholar
  4. 4.
    Suzuki A, Takashima S, Mizuguchi M, Kato M, Kunishita T, Tabira T (1994) Tohoku J Exp Med 174:181–187Google Scholar
  5. 5.
    Scheuner D, Eckman C, Jensen M, Song X, Citron M, Suzuki N, Bird TD, Hardy J, Hutton M, Kukull W, Larson E, Levy-Lahad E, Viitanen M, Peskind E, Poorkaj P, Schellenberg G, Tanzi R, Wasco W, Lannfelt L, Selkoe D, Younkin S (1996) Nat Med 2:864–870CrossRefPubMedGoogle Scholar
  6. 6.
    Hilbich C, Kisters-Woike B, Reed J, Masters CL, Beyreuther K (1991) J Mol Biol 218:149–163PubMedGoogle Scholar
  7. 7.
    Hilbich C, Kisters-Woike B, Reed J, Masters CL, Beyreuther K (1992) J Mol Biol 228:460–473PubMedGoogle Scholar
  8. 8.
    Barrow CJ, Zagorski MG (1991) Science 253:179–182PubMedGoogle Scholar
  9. 9.
    Barrow CJ, Yasuda A, Kenny PT, Zagorski MG (1992) J Mol Biol 225:1075–1093PubMedGoogle Scholar
  10. 10.
    Jarrett JT, Lansbury PT Jr (1993) Cell 73:1055–1058CrossRefPubMedGoogle Scholar
  11. 11.
    Jarrett JT, Berger EP, Lansbury PT Jr (1993) Biochemistry 32:4693–4697PubMedGoogle Scholar
  12. 12.
    Come JH, Fraser PE, Lansbury PT Jr (1993) Proc Natl Acad Sci USA 90:5959–5963PubMedGoogle Scholar
  13. 13.
    Bush AI, Pettingell WH Jr, Paradis MD, Tanzi RE (1994) J Biol Chem 269:12152–12158PubMedGoogle Scholar
  14. 14.
    Bush AI, Pettingell WH, Multhaup G, Paradis MD, Vonsattel J-P, Gusella JF, Beyreuther K, Masters CL, Tanzi RE (1994) Science 265:1464–1467PubMedGoogle Scholar
  15. 15.
    Garzon-Rodriguez W, Sepulveda-Becerra M, Milton S, Glabe CG (1997) J Biol Chem 272:21037–21044CrossRefPubMedGoogle Scholar
  16. 16.
    Huang X, Atwood CS, Moir RD, Hartshorn MA, Vonsattel JP, Tanzi RE, Bush AI (1997) J Biol Chem 272:26464–26470CrossRefPubMedGoogle Scholar
  17. 17.
    Atwood CS, Moir RD, Huang X, Scarpa RC, Bacarra NM, Romano DM, Hartshorn MA, Tanzi RE, Bush AI (1998) J Biol Chem 273:12817–12826CrossRefPubMedGoogle Scholar
  18. 18.
    Lovell MA, Robertson JD, Teesdale WJ, Campbell JL, Markesbery WR (1998) J Neurol Sci 158:47–52CrossRefPubMedGoogle Scholar
  19. 19.
    Lee JY, Mook-Jung I, Koh JY (1999) J Neurosci 19:RC10PubMedGoogle Scholar
  20. 20.
    Suh SW, Jensen KB, Jensen MS, Silva DS, Kesslak PJ, Danscher G, Frederickson CJ (2000) Brain Res 852:274–278CrossRefPubMedGoogle Scholar
  21. 21.
    Lee JY, Cole TB, Palmiter RD, Suh SW, Koh JY (2002) Proc Natl Acad Sci USA 99:7705–7710CrossRefPubMedGoogle Scholar
  22. 22.
    Friedlich AL, Lee JY, van Groen T, Cherny RA, Volitakis I, Cole TB, Palmiter RD, Koh JY, Bush AI (2004) J Neurosci 24:3453–3459CrossRefPubMedGoogle Scholar
  23. 23.
    Duff K, Eckman C, Zehr C, Yu X, Prada CM, Perez-tur J, Hutton M, Buee L, Harigaya Y, Yager D, Morgan D, Gordon MN, Holcomb L, Refolo L, Zenk B, Hardy J, Younkin S (1996) Nature 383:710–713CrossRefPubMedGoogle Scholar
  24. 24.
    Lambert MP, Barlow AK, Chromy BA, Edwards C, Freed R, Liosatos M, Morgan TE, Rozovsky I, Trommer B, Viola KL, Wals P, Zhang C, Finch CE, Krafft GA, Klein WL (1998) Proc Natl Acad Sci USA 95:6448–6453CrossRefPubMedGoogle Scholar
  25. 25.
    Roher AE, Chaney MO, Kuo YM, Webster SD, Stine WB, Haverkamp LJ, Woods AS, Cotter RJ, Tuohy JM, Krafft GA, Bonnell BS, Emmerling MR (1996) J Biol Chem 271:20631–20635CrossRefPubMedGoogle Scholar
  26. 26.
    Nakabayashi J, Yoshimura M, Morishima-Kawashima M, Funato H, Miyakawa T, Yamazaki T, Ihara Y (1998) J Neuropathol Exp Neurol 57:343–352PubMedGoogle Scholar
  27. 27.
    Enya M, Morishima-Kawashima M, Yoshimura M, Shinkai Y, Kusui K, Khan K, Games D, Schenk D, Sugihara S, Yamaguchi H, Ihara Y (1999) Am J Pathol 154:271–279PubMedGoogle Scholar
  28. 28.
    McLean CA, Cherny RA, Fraser FW, Fuller SJ, Smith MJ, Beyreuther K, Bush AI, Masters CL (1999) Ann Neurol 46:860–866Google Scholar
  29. 29.
    Walsh DM, Tseng BP, Rydel RE, Podlisny MB, Selkoe DJ (2000) Biochemistry 39:10831–10839CrossRefPubMedGoogle Scholar
  30. 30.
    Podlisny MB, Ostaszewski BL, Squazzo SL, Koo EH, Rydell RE, Teplow DB, Selkoe DJ (1995) J Biol Chem 270:9564–9570CrossRefPubMedGoogle Scholar
  31. 31.
    Podlisny MB, Walsh DM, Amarante P, Ostaszewski BL, Stimson ER, Maggio JE, Teplow DB, Selkoe DJ (1998) Biochemistry 37:3602–3611CrossRefPubMedGoogle Scholar
  32. 32.
    Xia W, Zhang J, Kholodenko D, Citron M, Podlisny MB, Teplow DB, Haass C, Seubert P, Koo EH, Selkoe DJ (1997) J Biol Chem 272:7977–7982CrossRefPubMedGoogle Scholar
  33. 33.
    Walsh DM, Klyubin I, Fadeeva JV, Cullen WK, Anwyl R, Wolfe MS, Rowan MJ, Selkoe DJ (2002) Nature 416:535–539CrossRefPubMedGoogle Scholar
  34. 34.
    Gong Y, Chang L, Viola KL, Lacor PN, Lambert MP, Finch CE, Krafft GA, Klein WL (2003) Proc Natl Acad Sci USA 100:10417–10422CrossRefPubMedGoogle Scholar
  35. 35.
    Kayed R, Head E, Thompson JL, McIntire TM, Milton SC, Cotman CW, Glabe CG (2003) Science 300:486–489CrossRefPubMedGoogle Scholar
  36. 36.
    Atwood CS, Perry G, Zeng H, Kato Y, Jones WD, Ling KQ, Huang X, Moir RD, Wang D, Sayre LM, Smith MA, Chen SG, Bush AI (2004) Biochemistry 43:560–568CrossRefPubMedGoogle Scholar
  37. 37.
    Huang X, Olmez I, Aras NK, Gordon GE (1994) Atmos Environ 28:1385–1391CrossRefGoogle Scholar
  38. 38.
    Wood SJ, Maleeff B, Hart T, Wetzel R (1996) J Mol Biol 256:870–877CrossRefPubMedGoogle Scholar
  39. 39.
    Klunk WE, Pettegrew JW, Abraham DJ (1989) J Histochem Cytochem 37:1293–1297PubMedGoogle Scholar
  40. 40.
    Bush AI, Moir RD, Rosenkranz KM, Tanzi RE (1995) Science 268:1921–1923Google Scholar
  41. 41.
    LeVine HD (1993) Protein Sci 2:404–410PubMedGoogle Scholar
  42. 42.
    Huang X, Atwood CS, Goldstein LE, Hartshorn MA, Saunders AJ, Moir RD, Tanzi RE, Bush AI (1999) In: Iqbal K, Swaab DF, Winblad B, Wisniewski HM (eds) Alzheimer’s disease and related disorders: etiology, pathogenesis and therapeutics. Wiley, Chichester, UK, pp 383–390Google Scholar
  43. 43.
    Huang X, Atwood CS, Hartshorn MA, Multhaup G, Goldstein LE, Scrapa SC, Cuajungco MP, Gray DN, Lim J, Moir RD, Tanzi RE, Bush AI (1999) Biochemistry 38:7609–7616CrossRefPubMedGoogle Scholar
  44. 44.
    Curtain CC, Ali F, Volitakis I, Cherny RA, Norton RS, Beyreuther K, Barrow CJ, Masters CL, Bush AI, Barnham KJ (2001) J Biol Chem 276: 20466–20473CrossRefPubMedGoogle Scholar
  45. 45.
    Sengupta P, Garai K, Sahoo B, Shi Y, Callaway DJ, Maiti S (2003) Biochemistry 42:10506–10513CrossRefPubMedGoogle Scholar
  46. 46.
    Loeffler DA, LeWitt PA, Juneau PL, Sima AA, Nguyen HU, DeMaggio AJ, Brickman CM, Brewer GJ, Dick RD, Troyer MD, Kanaley L (1996) Brain Res 738:265–274CrossRefPubMedGoogle Scholar
  47. 47.
    Van Gool D, De Strooper B, Van Leuven F, Triau E, Dom R (1993) Neurobiol Aging 14:233–237CrossRefPubMedGoogle Scholar
  48. 48.
    Grundke-Iqbal I, Fleming J, Tung YC, Lassmann H, Iqbal K, Joshi JG (1990) Acta Neuropathol (Berlin) 81:105–110Google Scholar
  49. 49.
    Loeffler DA, Connor JR, Juneau PL, Snyder BS, Kanaley L, DeMaggio AJ, Nguyen H, Brickman CM, LeWitt PA (1995) J Neurochem 65:710–724PubMedGoogle Scholar
  50. 50.
    Cherny RA, Atwood CS, Xilinas ME, Gray DN, Jones WD, McLean CA, Barnham KJ, Volitakis I, Fraser FW, Kim Y, Huang X, Goldstein LE, Moir RD, Lim JT, Beyreuther K, Zheng H, Tanzi RE, Masters CL, Bush AI (2001) Neuron 30:665–676CrossRefPubMedGoogle Scholar
  51. 51.
    Regland B, Lehmann W, Abedini I, Blennow K, Jonsson M, Karlsson I, Sjogren M, Wallin A, Xilinas M, Gottfries CG (2001) Dement Geriatr Cogn Disord 12:408–414CrossRefPubMedGoogle Scholar
  52. 52.
    Ritchie CW, Bush AI, Mackinnon A, Macfarlane S, Mastwyk M, MacGregor L, Kiers L, Cherny R, Li QX, Tammer A, Carrington D, Mavros C, Volitakis I, Xilinas M, Ames D, Davis S, Beyreuther K, Tanzi RE, Masters CL (2003) Arch Neurol 60:1685–1691CrossRefPubMedGoogle Scholar

Copyright information

© SBIC 2004

Authors and Affiliations

  • Xudong Huang
    • 1
  • Craig S. Atwood
    • 1
    • 3
  • Robert D. Moir
    • 2
  • Mariana A. Hartshorn
    • 1
  • Rudolph E. Tanzi
    • 2
  • Ashley I. Bush
    • 1
  1. 1.Department of Psychiatry, and Laboratory for Oxidation Biology, Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative DiseaseMassachusetts General Hospital and Harvard Medical SchoolBostonUSA
  2. 2.Department of Neurology, Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative DiseaseMassachusetts General Hospital and Harvard Medical SchoolBostonUSA
  3. 3.Department of MedicineUniversity of WisconsinMadisonUSA

Personalised recommendations