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. 1997 Apr 15;323(Pt 2):539–546. doi: 10.1042/bj3230539

Binding of Abeta to alpha- and beta-synucleins: identification of segments in alpha-synuclein/NAC precursor that bind Abeta and NAC.

P H Jensen 1, P Hojrup 1, H Hager 1, M S Nielsen 1, L Jacobsen 1, O F Olesen 1, J Gliemann 1, R Jakes 1
PMCID: PMC1218353  PMID: 9163350

Abstract

NAC, a 35-residue peptide derived from the neuronal protein alpha-synuclein/NAC precursor, is tightly associated with Abeta fibrils in Alzheimer's disease amyloid, and alpha-synuclein has recently been shown to bind Abeta in vitro. We have studied the interaction between Abeta and synucleins, aiming at determining segments in alpha-synuclein that can account for the binding, as well as identifying a possible interaction between Abeta and the beta-type synuclein. We report that Abeta binds to native and recombinant alpha-synuclein, and to beta-synuclein in an SDS-sensitive interaction (IC50 approx. 20 microM), as determined by chemical cross-linking and solid-phase binding assays. alpha-Synuclein and beta-synuclein were found to stimulate Abeta-aggregation in vitro to the same extent. The synucleins also displayed Abeta-inhibitable binding of NAC and they were capable of forming dimers. Using proteolytic fragmentation of alpha-synuclein and cross-linking to 125I-Abeta, we identified two consecutive binding domains (residues 1-56 and 57-97) by Edman degradation and mass spectrometric analysis, and a synthetic peptide comprising residues 32-57 possessed Abeta-binding activity. To test further the possible significance in pathology, alpha-synuclein was biotinylated and shown to bind specifically to amyloid plaques in a brain with Alzheimer's disease. It is proposed that the multiple Abeta-binding sites in alpha-synuclein are involved in the development of amyloid plaques.

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Selected References

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  1. Eriksson S., Janciauskiene S., Lannfelt L. Alpha 1-antichymotrypsin regulates Alzheimer beta-amyloid peptide fibril formation. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2313–2317. doi: 10.1073/pnas.92.6.2313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fraser P. E., Nguyen J. T., McLachlan D. R., Abraham C. R., Kirschner D. A. Alpha 1-antichymotrypsin binding to Alzheimer A beta peptides is sequence specific and induces fibril disaggregation in vitro. J Neurochem. 1993 Jul;61(1):298–305. doi: 10.1111/j.1471-4159.1993.tb03568.x. [DOI] [PubMed] [Google Scholar]
  3. Games D., Adams D., Alessandrini R., Barbour R., Berthelette P., Blackwell C., Carr T., Clemens J., Donaldson T., Gillespie F. Alzheimer-type neuropathology in transgenic mice overexpressing V717F beta-amyloid precursor protein. Nature. 1995 Feb 9;373(6514):523–527. doi: 10.1038/373523a0. [DOI] [PubMed] [Google Scholar]
  4. George J. M., Jin H., Woods W. S., Clayton D. F. Characterization of a novel protein regulated during the critical period for song learning in the zebra finch. Neuron. 1995 Aug;15(2):361–372. doi: 10.1016/0896-6273(95)90040-3. [DOI] [PubMed] [Google Scholar]
  5. Golabek A., Marques M. A., Lalowski M., Wisniewski T. Amyloid beta binding proteins in vitro and in normal human cerebrospinal fluid. Neurosci Lett. 1995 May 19;191(1-2):79–82. doi: 10.1016/0304-3940(95)11565-7. [DOI] [PubMed] [Google Scholar]
  6. Han H., Weinreb P. H., Lansbury P. T., Jr The core Alzheimer's peptide NAC forms amyloid fibrils which seed and are seeded by beta-amyloid: is NAC a common trigger or target in neurodegenerative disease? Chem Biol. 1995 Mar;2(3):163–169. doi: 10.1016/1074-5521(95)90071-3. [DOI] [PubMed] [Google Scholar]
  7. Iwai A., Masliah E., Yoshimoto M., Ge N., Flanagan L., de Silva H. A., Kittel A., Saitoh T. The precursor protein of non-A beta component of Alzheimer's disease amyloid is a presynaptic protein of the central nervous system. Neuron. 1995 Feb;14(2):467–475. doi: 10.1016/0896-6273(95)90302-x. [DOI] [PubMed] [Google Scholar]
  8. Iwai A., Yoshimoto M., Masliah E., Saitoh T. Non-A beta component of Alzheimer's disease amyloid (NAC) is amyloidogenic. Biochemistry. 1995 Aug 15;34(32):10139–10145. doi: 10.1021/bi00032a006. [DOI] [PubMed] [Google Scholar]
  9. Jakes R., Spillantini M. G., Goedert M. Identification of two distinct synucleins from human brain. FEBS Lett. 1994 May 23;345(1):27–32. doi: 10.1016/0014-5793(94)00395-5. [DOI] [PubMed] [Google Scholar]
  10. Jensen P. H., Sørensen E. S., Petersen T. E., Gliemann J., Rasmussen L. K. Residues in the synuclein consensus motif of the alpha-synuclein fragment, NAC, participate in transglutaminase-catalysed cross-linking to Alzheimer-disease amyloid beta A4 peptide. Biochem J. 1995 Aug 15;310(Pt 1):91–94. doi: 10.1042/bj3100091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jiang H., Burdick D., Glabe C. G., Cotman C. W., Tenner A. J. beta-Amyloid activates complement by binding to a specific region of the collagen-like domain of the C1q A chain. J Immunol. 1994 May 15;152(10):5050–5059. [PubMed] [Google Scholar]
  12. Ma J., Yee A., Brewer H. B., Jr, Das S., Potter H. Amyloid-associated proteins alpha 1-antichymotrypsin and apolipoprotein E promote assembly of Alzheimer beta-protein into filaments. Nature. 1994 Nov 3;372(6501):92–94. doi: 10.1038/372092a0. [DOI] [PubMed] [Google Scholar]
  13. Maroteaux L., Campanelli J. T., Scheller R. H. Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J Neurosci. 1988 Aug;8(8):2804–2815. doi: 10.1523/JNEUROSCI.08-08-02804.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Olesen O. F., Mikkelsen J. D., Gerdes C., Jensen P. H. Isoform-specific binding of human apolipoprotein E to the non-amyloid beta component of Alzheimer's disease amyloid. Brain Res Mol Brain Res. 1997 Feb;44(1):105–112. doi: 10.1016/s0169-328x(96)00196-9. [DOI] [PubMed] [Google Scholar]
  15. Rasmussen L. K., Sørensen E. S., Petersen T. E., Gliemann J., Jensen P. H. Identification of glutamine and lysine residues in Alzheimer amyloid beta A4 peptide responsible for transglutaminase-catalysed homopolymerization and cross-linking to alpha 2M receptor. FEBS Lett. 1994 Jan 31;338(2):161–166. doi: 10.1016/0014-5793(94)80356-0. [DOI] [PubMed] [Google Scholar]
  16. Rogers J., Cooper N. R., Webster S., Schultz J., McGeer P. L., Styren S. D., Civin W. H., Brachova L., Bradt B., Ward P. Complement activation by beta-amyloid in Alzheimer disease. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10016–10020. doi: 10.1073/pnas.89.21.10016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schellenberg G. D. Genetic dissection of Alzheimer disease, a heterogeneous disorder. Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8552–8559. doi: 10.1073/pnas.92.19.8552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  19. Selkoe D. J. Normal and abnormal biology of the beta-amyloid precursor protein. Annu Rev Neurosci. 1994;17:489–517. doi: 10.1146/annurev.ne.17.030194.002421. [DOI] [PubMed] [Google Scholar]
  20. Shibayama-Imazu T., Okahashi I., Omata K., Nakajo S., Ochiai H., Nakai Y., Hama T., Nakamura Y., Nakaya K. Cell and tissue distribution and developmental change of neuron specific 14 kDa protein (phosphoneuroprotein 14). Brain Res. 1993 Sep 17;622(1-2):17–25. doi: 10.1016/0006-8993(93)90796-p. [DOI] [PubMed] [Google Scholar]
  21. Strittmatter W. J., Weisgraber K. H., Huang D. Y., Dong L. M., Salvesen G. S., Pericak-Vance M., Schmechel D., Saunders A. M., Goldgaber D., Roses A. D. Binding of human apolipoprotein E to synthetic amyloid beta peptide: isoform-specific effects and implications for late-onset Alzheimer disease. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8098–8102. doi: 10.1073/pnas.90.17.8098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Strittmatter W. J., Weisgraber K. H., Huang D. Y., Dong L. M., Salvesen G. S., Pericak-Vance M., Schmechel D., Saunders A. M., Goldgaber D., Roses A. D. Binding of human apolipoprotein E to synthetic amyloid beta peptide: isoform-specific effects and implications for late-onset Alzheimer disease. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8098–8102. doi: 10.1073/pnas.90.17.8098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tjernberg L. O., Näslund J., Lindqvist F., Johansson J., Karlström A. R., Thyberg J., Terenius L., Nordstedt C. Arrest of beta-amyloid fibril formation by a pentapeptide ligand. J Biol Chem. 1996 Apr 12;271(15):8545–8548. doi: 10.1074/jbc.271.15.8545. [DOI] [PubMed] [Google Scholar]
  24. Uéda K., Fukushima H., Masliah E., Xia Y., Iwai A., Yoshimoto M., Otero D. A., Kondo J., Ihara Y., Saitoh T. Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11282–11286. doi: 10.1073/pnas.90.23.11282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yoshimoto M., Iwai A., Kang D., Otero D. A., Xia Y., Saitoh T. NACP, the precursor protein of the non-amyloid beta/A4 protein (A beta) component of Alzheimer disease amyloid, binds A beta and stimulates A beta aggregation. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9141–9145. doi: 10.1073/pnas.92.20.9141. [DOI] [PMC free article] [PubMed] [Google Scholar]

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