Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 2000 Jun;78(6):2892–2899. doi: 10.1016/s0006-3495(00)76830-5

Poly-L-glutamine forms cation channels: relevance to the pathogenesis of the polyglutamine diseases.

H Monoi 1, S Futaki 1, S Kugimiya 1, H Minakata 1, K Yoshihara 1
PMCID: PMC1300875  PMID: 10827970

Abstract

We report that long-chain poly-L-glutamine forms cation-selective channels when incorporated into artificial planar lipid bilayer membranes. The channel was permeable to alkali cations and H(+) ions and virtually impermeable to anions; the selectivity sequence based on the single-channel conductance was H(+) >> Cs(+) > K(+) > Na(+). The cation channel was characterized by long-lived open states (often lasting for several minutes to tens of minutes) interrupted by brief closings. The appearance of the channel depended critically on the length of polyglutamine chains; ion channels were observed with 40-residue stretches, whereas no significant conductance changes were detected with 29-residue tracts. The channel-forming threshold length of poly-L-glutamine was thus between 29 and 40 residues. A molecular mechanics calculation suggests a mu-helix (. Biophys. J. 69:1130-1141) as a candidate molecular structure of the channel. The channel-forming nature of long-chain poly-L-glutamine may provide a clue to the elucidation of the pathogenetic mechanism of the polyglutamine diseases, a group of inherited neurodegenerative disorders including Huntington's disease.

Full Text

The Full Text of this article is available as a PDF (483.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrew S. E., Goldberg Y. P., Kremer B., Telenius H., Theilmann J., Adam S., Starr E., Squitieri F., Lin B., Kalchman M. A. The relationship between trinucleotide (CAG) repeat length and clinical features of Huntington's disease. Nat Genet. 1993 Aug;4(4):398–403. doi: 10.1038/ng0893-398. [DOI] [PubMed] [Google Scholar]
  2. Arispe N., Pollard H. B., Rojas E. Giant multilevel cation channels formed by Alzheimer disease amyloid beta-protein [A beta P-(1-40)] in bilayer membranes. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10573–10577. doi: 10.1073/pnas.90.22.10573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arispe N., Rojas E., Pollard H. B. Alzheimer disease amyloid beta protein forms calcium channels in bilayer membranes: blockade by tromethamine and aluminum. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):567–571. doi: 10.1073/pnas.90.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Banfi S., Servadio A., Chung M. Y., Kwiatkowski T. J., Jr, McCall A. E., Duvick L. A., Shen Y., Roth E. J., Orr H. T., Zoghbi H. Y. Identification and characterization of the gene causing type 1 spinocerebellar ataxia. Nat Genet. 1994 Aug;7(4):513–520. doi: 10.1038/ng0894-513. [DOI] [PubMed] [Google Scholar]
  5. Barron L. H., Warner J. P., Porteous M., Holloway S., Simpson S., Davidson R., Brock D. J. A study of the Huntington's disease associated trinucleotide repeat in the Scottish population. J Med Genet. 1993 Dec;30(12):1003–1007. doi: 10.1136/jmg.30.12.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper A. J., Sheu K. F., Burke J. R., Onodera O., Strittmatter W. J., Roses A. D., Blass J. P. Polyglutamine domains are substrates of tissue transglutaminase: does transglutaminase play a role in expanded CAG/poly-Q neurodegenerative diseases? J Neurochem. 1997 Jul;69(1):431–434. doi: 10.1046/j.1471-4159.1997.69010431.x. [DOI] [PubMed] [Google Scholar]
  7. David G., Abbas N., Stevanin G., Dürr A., Yvert G., Cancel G., Weber C., Imbert G., Saudou F., Antoniou E. Cloning of the SCA7 gene reveals a highly unstable CAG repeat expansion. Nat Genet. 1997 Sep;17(1):65–70. doi: 10.1038/ng0997-65. [DOI] [PubMed] [Google Scholar]
  8. DiFiglia M., Sapp E., Chase K. O., Davies S. W., Bates G. P., Vonsattel J. P., Aronin N. Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science. 1997 Sep 26;277(5334):1990–1993. doi: 10.1126/science.277.5334.1990. [DOI] [PubMed] [Google Scholar]
  9. Duyao M., Ambrose C., Myers R., Novelletto A., Persichetti F., Frontali M., Folstein S., Ross C., Franz M., Abbott M. Trinucleotide repeat length instability and age of onset in Huntington's disease. Nat Genet. 1993 Aug;4(4):387–392. doi: 10.1038/ng0893-387. [DOI] [PubMed] [Google Scholar]
  10. Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O., Tagliavini F. Neurotoxicity of a prion protein fragment. Nature. 1993 Apr 8;362(6420):543–546. doi: 10.1038/362543a0. [DOI] [PubMed] [Google Scholar]
  11. Gentile V., Sepe C., Calvani M., Melone M. A., Cotrufo R., Cooper A. J., Blass J. P., Peluso G. Tissue transglutaminase-catalyzed formation of high-molecular-weight aggregates in vitro is favored with long polyglutamine domains: a possible mechanism contributing to CAG-triplet diseases. Arch Biochem Biophys. 1998 Apr 15;352(2):314–321. doi: 10.1006/abbi.1998.0592. [DOI] [PubMed] [Google Scholar]
  12. Green H. Human genetic diseases due to codon reiteration: relationship to an evolutionary mechanism. Cell. 1993 Sep 24;74(6):955–956. doi: 10.1016/0092-8674(93)90718-6. [DOI] [PubMed] [Google Scholar]
  13. Ikeda H., Yamaguchi M., Sugai S., Aze Y., Narumiya S., Kakizuka A. Expanded polyglutamine in the Machado-Joseph disease protein induces cell death in vitro and in vivo. Nat Genet. 1996 Jun;13(2):196–202. doi: 10.1038/ng0696-196. [DOI] [PubMed] [Google Scholar]
  14. Imbert G., Saudou F., Yvert G., Devys D., Trottier Y., Garnier J. M., Weber C., Mandel J. L., Cancel G., Abbas N. Cloning of the gene for spinocerebellar ataxia 2 reveals a locus with high sensitivity to expanded CAG/glutamine repeats. Nat Genet. 1996 Nov;14(3):285–291. doi: 10.1038/ng1196-285. [DOI] [PubMed] [Google Scholar]
  15. Kahlem P., Green H., Djian P. Transglutaminase action imitates Huntington's disease: selective polymerization of Huntingtin containing expanded polyglutamine. Mol Cell. 1998 Mar;1(4):595–601. doi: 10.1016/s1097-2765(00)80059-3. [DOI] [PubMed] [Google Scholar]
  16. Kahlem P., Terré C., Green H., Djian P. Peptides containing glutamine repeats as substrates for transglutaminase-catalyzed cross-linking: relevance to diseases of the nervous system. Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14580–14585. doi: 10.1073/pnas.93.25.14580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kawaguchi Y., Okamoto T., Taniwaki M., Aizawa M., Inoue M., Katayama S., Kawakami H., Nakamura S., Nishimura M., Akiguchi I. CAG expansions in a novel gene for Machado-Joseph disease at chromosome 14q32.1. Nat Genet. 1994 Nov;8(3):221–228. doi: 10.1038/ng1194-221. [DOI] [PubMed] [Google Scholar]
  18. Koide R., Ikeuchi T., Onodera O., Tanaka H., Igarashi S., Endo K., Takahashi H., Kondo R., Ishikawa A., Hayashi T. Unstable expansion of CAG repeat in hereditary dentatorubral-pallidoluysian atrophy (DRPLA). Nat Genet. 1994 Jan;6(1):9–13. doi: 10.1038/ng0194-9. [DOI] [PubMed] [Google Scholar]
  19. Komure O., Sano A., Nishino N., Yamauchi N., Ueno S., Kondoh K., Sano N., Takahashi M., Murayama N., Kondo I. DNA analysis in hereditary dentatorubral-pallidoluysian atrophy: correlation between CAG repeat length and phenotypic variation and the molecular basis of anticipation. Neurology. 1995 Jan;45(1):143–149. doi: 10.1212/wnl.45.1.143. [DOI] [PubMed] [Google Scholar]
  20. Kunz B., Sandmeier E., Christen P. Neurotoxicity of prion peptide 106-126 not confirmed. FEBS Lett. 1999 Sep 10;458(1):65–68. doi: 10.1016/s0014-5793(99)01123-0. [DOI] [PubMed] [Google Scholar]
  21. La Spada A. R., Wilson E. M., Lubahn D. B., Harding A. E., Fischbeck K. H. Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy. Nature. 1991 Jul 4;352(6330):77–79. doi: 10.1038/352077a0. [DOI] [PubMed] [Google Scholar]
  22. Li S. H., Schilling G., Young W. S., 3rd, Li X. J., Margolis R. L., Stine O. C., Wagster M. V., Abbott M. H., Franz M. L., Ranen N. G. Huntington's disease gene (IT15) is widely expressed in human and rat tissues. Neuron. 1993 Nov;11(5):985–993. doi: 10.1016/0896-6273(93)90127-d. [DOI] [PubMed] [Google Scholar]
  23. Lin M. C., Mirzabekov T., Kagan B. L. Channel formation by a neurotoxic prion protein fragment. J Biol Chem. 1997 Jan 3;272(1):44–47. doi: 10.1074/jbc.272.1.44. [DOI] [PubMed] [Google Scholar]
  24. Monoi H. New tubular single-stranded helix of poly-L-amino acids suggested by molecular mechanics calculations: I. Homopolypeptides in isolated environments. Biophys J. 1995 Sep;69(3):1130–1141. doi: 10.1016/S0006-3495(95)79987-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nagafuchi S., Yanagisawa H., Ohsaki E., Shirayama T., Tadokoro K., Inoue T., Yamada M. Structure and expression of the gene responsible for the triplet repeat disorder, dentatorubral and pallidoluysian atrophy (DRPLA). Nat Genet. 1994 Oct;8(2):177–182. doi: 10.1038/ng1094-177. [DOI] [PubMed] [Google Scholar]
  26. Nagafuchi S., Yanagisawa H., Sato K., Shirayama T., Ohsaki E., Bundo M., Takeda T., Tadokoro K., Kondo I., Murayama N. Dentatorubral and pallidoluysian atrophy expansion of an unstable CAG trinucleotide on chromosome 12p. Nat Genet. 1994 Jan;6(1):14–18. doi: 10.1038/ng0194-14. [DOI] [PubMed] [Google Scholar]
  27. Novelletto A., Persichetti F., Sabbadini G., Mandich P., Bellone E., Ajmar F., Pergola M., Del Senno L., MacDonald M. E., Gusella J. F. Analysis of the trinucleotide repeat expansion in Italian families affected with Huntington disease. Hum Mol Genet. 1994 Jan;3(1):93–98. doi: 10.1093/hmg/3.1.93. [DOI] [PubMed] [Google Scholar]
  28. Orr H. T., Chung M. Y., Banfi S., Kwiatkowski T. J., Jr, Servadio A., Beaudet A. L., McCall A. E., Duvick L. A., Ranum L. P., Zoghbi H. Y. Expansion of an unstable trinucleotide CAG repeat in spinocerebellar ataxia type 1. Nat Genet. 1993 Jul;4(3):221–226. doi: 10.1038/ng0793-221. [DOI] [PubMed] [Google Scholar]
  29. Paulson H. L., Perez M. K., Trottier Y., Trojanowski J. Q., Subramony S. H., Das S. S., Vig P., Mandel J. L., Fischbeck K. H., Pittman R. N. Intranuclear inclusions of expanded polyglutamine protein in spinocerebellar ataxia type 3. Neuron. 1997 Aug;19(2):333–344. doi: 10.1016/s0896-6273(00)80943-5. [DOI] [PubMed] [Google Scholar]
  30. Perutz M. F., Johnson T., Suzuki M., Finch J. T. Glutamine repeats as polar zippers: their possible role in inherited neurodegenerative diseases. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5355–5358. doi: 10.1073/pnas.91.12.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ranum L. P., Chung M. Y., Banfi S., Bryer A., Schut L. J., Ramesar R., Duvick L. A., McCall A., Subramony S. H., Goldfarb L. Molecular and clinical correlations in spinocerebellar ataxia type I: evidence for familial effects on the age at onset. Am J Hum Genet. 1994 Aug;55(2):244–252. [PMC free article] [PubMed] [Google Scholar]
  32. Roseman M. A. Hydrophobicity of the peptide C=O...H-N hydrogen-bonded group. J Mol Biol. 1988 Jun 5;201(3):621–623. doi: 10.1016/0022-2836(88)90642-0. [DOI] [PubMed] [Google Scholar]
  33. Rubinsztein D. C., Leggo J., Coles R., Almqvist E., Biancalana V., Cassiman J. J., Chotai K., Connarty M., Crauford D., Curtis A. Phenotypic characterization of individuals with 30-40 CAG repeats in the Huntington disease (HD) gene reveals HD cases with 36 repeats and apparently normal elderly individuals with 36-39 repeats. Am J Hum Genet. 1996 Jul;59(1):16–22. [PMC free article] [PubMed] [Google Scholar]
  34. Sanpei K., Takano H., Igarashi S., Sato T., Oyake M., Sasaki H., Wakisaka A., Tashiro K., Ishida Y., Ikeuchi T. Identification of the spinocerebellar ataxia type 2 gene using a direct identification of repeat expansion and cloning technique, DIRECT. Nat Genet. 1996 Nov;14(3):277–284. doi: 10.1038/ng1196-277. [DOI] [PubMed] [Google Scholar]
  35. Servadio A., Koshy B., Armstrong D., Antalffy B., Orr H. T., Zoghbi H. Y. Expression analysis of the ataxin-1 protein in tissues from normal and spinocerebellar ataxia type 1 individuals. Nat Genet. 1995 May;10(1):94–98. doi: 10.1038/ng0595-94. [DOI] [PubMed] [Google Scholar]
  36. Sharp A. H., Loev S. J., Schilling G., Li S. H., Li X. J., Bao J., Wagster M. V., Kotzuk J. A., Steiner J. P., Lo A. Widespread expression of Huntington's disease gene (IT15) protein product. Neuron. 1995 May;14(5):1065–1074. doi: 10.1016/0896-6273(95)90345-3. [DOI] [PubMed] [Google Scholar]
  37. Snell R. G., MacMillan J. C., Cheadle J. P., Fenton I., Lazarou L. P., Davies P., MacDonald M. E., Gusella J. F., Harper P. S., Shaw D. J. Relationship between trinucleotide repeat expansion and phenotypic variation in Huntington's disease. Nat Genet. 1993 Aug;4(4):393–397. doi: 10.1038/ng0893-393. [DOI] [PubMed] [Google Scholar]
  38. Stine O. C., Pleasant N., Franz M. L., Abbott M. H., Folstein S. E., Ross C. A. Correlation between the onset age of Huntington's disease and length of the trinucleotide repeat in IT-15. Hum Mol Genet. 1993 Oct;2(10):1547–1549. doi: 10.1093/hmg/2.10.1547. [DOI] [PubMed] [Google Scholar]
  39. Stott K., Blackburn J. M., Butler P. J., Perutz M. Incorporation of glutamine repeats makes protein oligomerize: implications for neurodegenerative diseases. Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6509–6513. doi: 10.1073/pnas.92.14.6509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Strong T. V., Tagle D. A., Valdes J. M., Elmer L. W., Boehm K., Swaroop M., Kaatz K. W., Collins F. S., Albin R. L. Widespread expression of the human and rat Huntington's disease gene in brain and nonneural tissues. Nat Genet. 1993 Nov;5(3):259–265. doi: 10.1038/ng1193-259. [DOI] [PubMed] [Google Scholar]
  41. Trottier Y., Devys D., Imbert G., Saudou F., An I., Lutz Y., Weber C., Agid Y., Hirsch E. C., Mandel J. L. Cellular localization of the Huntington's disease protein and discrimination of the normal and mutated form. Nat Genet. 1995 May;10(1):104–110. doi: 10.1038/ng0595-104. [DOI] [PubMed] [Google Scholar]
  42. Yazawa I., Nukina N., Hashida H., Goto J., Yamada M., Kanazawa I. Abnormal gene product identified in hereditary dentatorubral-pallidoluysian atrophy (DRPLA) brain. Nat Genet. 1995 May;10(1):99–103. doi: 10.1038/ng0595-99. [DOI] [PubMed] [Google Scholar]
  43. Zoghbi H. Y., Orr H. T. Polyglutamine diseases: protein cleavage and aggregation. Curr Opin Neurobiol. 1999 Oct;9(5):566–570. doi: 10.1016/S0959-4388(99)00013-6. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES