1993 Fiscal Year Annual Research Report

アルツハイマ-神経原線維変化(PHF)内tauの翻訳後修飾の同定

Research Project

Project/Area Number	03102008
Research Institution	University of Tokyo
Principal Investigator	井原康夫東京大学, 医学部(医), 教授 (60114386)
Keywords	アルツハイマ-病 / PHF / tau / リン酸化 / proline-directed protein kinase / ubiquitin
Research Abstract	アルツハイマ-神経原線維変化(PHF)の翻訳後修飾にはリン酸化、ユビキチン化、およびcrosslinkingが知られている。本研究では、前2者に関して明確にした。 1リン酸化 (1)胎児型tauのリン酸化部位の同定リン酸化依存性のPHF polyclonalおよびmonoclonalが胎児型tauを認識することからPHF-tauとfetal tauのリン酸化に共通性があることが明らかとなった。PHF-tauのリン酸化部位の同定にはいくつかの困難があったので(後述参照)、PHF-tauとリン酸化において類似性を示すfetal tauのリン酸化部位の決定をした。リン酸化部位の同定にはイオンスプレ-型質量分析機、エタンチオ-ル修飾後のアミノ酸配列分析を用いた。その結果、(ヒト441残基イソフォ-ムを基準として)、Thr-181,Ser-198,-199,-202,Thr-231,Ser-235,-396,-400,-404,-413がリン酸化部位として同定された。Ser-198,-400,-413を除いてすべてSer/Thr-Proのmotifでありプロリン指向性リン酸化といえこれらのリン酸化部位はTPKII/cdk5およびTPKI/GSK3βによるリン酸化部位を合わせたものに大体致した。 (2)PHF-tauのリン酸化部位の同定 PHF-tauのリン酸化部位の同定は困難をきわめた。第1にPHF-tauがプロテア-ゼに非常に耐性であること、第2にリン酸化ペプチドの逆相カラムからの収率が非常に悪いこと、が大きな要因であった。試行錯誤の結果、上記と同様な方法を用いて、Ser-198,-199,-202,-205,-208,-210,Thr-212,Ser-214,Thr-217,Ser-231,-235,-262,-396,-400,-403,-404,-409,-412,-413,-422が異常ン酸化部位として同定された。以上のようにPHF-tauはfetal tauよりはるかにリン酸化されていた

Research Products
(26 results)

All Other

All Publications (26 results)

[Publications] Yamaguchi H et al.: "Amyloid β/A4 protein precursor is bound to neurofibrillary tangles in Alzheimer-type dementia." Brain Res. 537. 318-322 (1991)
[Publications] Yamaguchi H et al.: "Secondary deposition of β amyloid within extracellular neurofibrillary tangles in Alzheimer-type dementia." Am.J.Pathol.138. 699-705 (1991)
[Publications] Yamaguchi H et al.: "Extracellular neurofibrillary tangles associated with de generating neurites and neuropil threads inAlzheimer-type dementia." Acta Neuropathol.81. 603-609 (1991)
[Publications] Oyama F et al.: "Differential expression of β amyloid protein precursor(APP)and tau mRNA in the aged human brain:Individual variability and correlation between APP-751 and four-repeat tau." J.Neuropathol.Exp.Neurol.50. 560-578 (1991)
[Publications] Morishima M et al.: "A possible fetal antigen of Mr 70,000 in neurofibrillary tangles." Brain Res.554. 316-320 (1991)
[Publications] Uchida Y et al.: "The growth in-hibitory factor that is deficient in the Alzheimer's disease brain is a 68 amino acid metallothionein like protein." Neuron. 7. 337-347 (1991)
[Publications] Ihara Y et al.: "The regenerative process of Alzheimer's disease.Proceedings of the XIth International Congress of Neuropathology." Neuropathology. 4. 110-115 (1991)
[Publications] Ivy Go et al.: "The protease inhibitor leupeptin induces several signs of aging in brain,retina and internal organs of young rats." Arch Gerontol Geriat. 12. 119-131 (1991)
[Publications] Tokuda T et al.: "Re-examination of ex-boxers' brains using immunohistochemistry with antibodies to amyloid beta protein and tau protein." Acta Neuropathol. 82. 280-285 (1991)
[Publications] Mizutani T et al.: "Clinicopathologic study of dementia in Parkinson disease." Dementia. 2. 229-236 (1991)
[Publications] Watanabe N et al.: "Tau 2:A probe for a Ser-conformation in the amino terminus of τ." J Neurochem. 58. 960-966 (1992)
[Publications] Kanemaru K et al.: "Fetal-type phosphorylation of the tau in paired helical filaments." J Neurochem. 58. 1667-1675 (1992)
[Publications] Choi-Miura NH et al.: "SP40,40 is a constituent of Alzheimer's amyloid." Acta Neuropathol. 83. 260-264 (1992)
[Publications] Iwatsubo T et al.: "Lack of ubiquitin immunoreactivities at both ends of neuropil threads-possible bidirectional growth of neuropil threads-." Am J Pathol. 140. 277-282 (1992)
[Publications] Hasegawa M et al.: "Protein sequence and mass spectrometric analyses of tau in the Alzheimer's disease" J Biol Chem. 267. 17047-17054 (1992)
[Publications] Oyama F et al.: "A novel correlation between the levels of β-amyloid protein precursor and tau transcripts in the aged human brain." J Neurochem. 59. 1117-1125 (1992)
[Publications] Yanagisawa K et al.: "Secretory pathway of β/A4 amyloid protein precursor in familial Alzheimer's disease with Val_<717> to Ile mutation." Neurosci Lett. 144. 43-45 (1992)
[Publications] Tsuji S et al.: "Molecular cloning ofhuman growth inhibitory factor cDNA and its down-regulation in Alzheimer's disease." EMBO J. 11. 4843-4850 (1992)
[Publications] Oyama F et al.: "β-Amyloid protein precursor and tau mRNA levels versus β-amyloid plaque and neurofibrillary tangles in the aged human brain." J Neurochem. 60. 1658-1664 (1993)
[Publications] Hasegawa M et al.: "Characterization of two distinct monoclonal antibodies to paired helical filaments(PHF)-Further evidence for fetal-type phosphorylation of the tau in PHF-." J Neurochem. 60. 2068-2077 (1993)
[Publications] Morishima M et al.: "Ubiquitin is conjugated with amino-terminally processed tau in paired helical filaments." Neuron. 10. 1151-1160 (1993)
[Publications] Yoshida H et al.: "tau in paired helical filaments(PHF)is functionally distinct from fetal tau.-assembly incompetence of PHF-tau-." J Neurochem. 61. 1183-1186 (1993)
[Publications] Kobayashi H et al.: "Molecular cloning of rat growth inhibitory factor cDNA and the expression in the central nervous system." Mol Brain Res. 19. 188-194 (1993)
[Publications] Watanabe A et al.: "In vivo phosphorylation sites in fetal and adult rat tau." J Biol Chem. 268. 25712-25717 (1993)
[Publications] Ueda K et al.: "Molecular cloning of a novel component of amyloid in Alzheimer's disease." Proc Natl Acad Sci USA. 90. 11282-11286 (1993)
[Publications] Oyama F et al: "Down's syndrome:Upregulation of β- amyloid protien precursor and tau mRNAs and their defective coordinatioin." J Neurochem. (in press).

1993 Fiscal Year Annual Research Report

アルツハイマ-神経原線維変化(PHF)内tauの翻訳後修飾の同定

Principal Investigator

井原 康夫 東京大学, 医学部(医), 教授 (60114386)

Research Products

[Publications] Yamaguchi H et al.: "Amyloid β/A4 protein precursor is bound to neurofibrillary tangles in Alzheimer-type dementia." Brain Res. 537. 318-322 (1991)

[Publications] Yamaguchi H et al.: "Secondary deposition of β amyloid within extracellular neurofibrillary tangles in Alzheimer-type dementia." Am.J.Pathol.138. 699-705 (1991)

[Publications] Yamaguchi H et al.: "Extracellular neurofibrillary tangles associated with de generating neurites and neuropil threads inAlzheimer-type dementia." Acta Neuropathol.81. 603-609 (1991)

[Publications] Oyama F et al.: "Differential expression of β amyloid protein precursor(APP)and tau mRNA in the aged human brain:Individual variability and correlation between APP-751 and four-repeat tau." J.Neuropathol.Exp.Neurol.50. 560-578 (1991)

[Publications] Morishima M et al.: "A possible fetal antigen of Mr 70,000 in neurofibrillary tangles." Brain Res.554. 316-320 (1991)

[Publications] Uchida Y et al.: "The growth in-hibitory factor that is deficient in the Alzheimer's disease brain is a 68 amino acid metallothionein like protein." Neuron. 7. 337-347 (1991)

[Publications] Ihara Y et al.: "The regenerative process of Alzheimer's disease.Proceedings of the XIth International Congress of Neuropathology." Neuropathology. 4. 110-115 (1991)

[Publications] Ivy Go et al.: "The protease inhibitor leupeptin induces several signs of aging in brain,retina and internal organs of young rats." Arch Gerontol Geriat. 12. 119-131 (1991)

[Publications] Tokuda T et al.: "Re-examination of ex-boxers' brains using immunohistochemistry with antibodies to amyloid beta protein and tau protein." Acta Neuropathol. 82. 280-285 (1991)

[Publications] Mizutani T et al.: "Clinicopathologic study of dementia in Parkinson disease." Dementia. 2. 229-236 (1991)

[Publications] Watanabe N et al.: "Tau 2:A probe for a Ser-conformation in the amino terminus of τ." J Neurochem. 58. 960-966 (1992)

[Publications] Kanemaru K et al.: "Fetal-type phosphorylation of the tau in paired helical filaments." J Neurochem. 58. 1667-1675 (1992)

[Publications] Choi-Miura NH et al.: "SP40,40 is a constituent of Alzheimer's amyloid." Acta Neuropathol. 83. 260-264 (1992)

[Publications] Iwatsubo T et al.: "Lack of ubiquitin immunoreactivities at both ends of neuropil threads-possible bidirectional growth of neuropil threads-." Am J Pathol. 140. 277-282 (1992)

[Publications] Hasegawa M et al.: "Protein sequence and mass spectrometric analyses of tau in the Alzheimer's disease" J Biol Chem. 267. 17047-17054 (1992)

[Publications] Oyama F et al.: "A novel correlation between the levels of β-amyloid protein precursor and tau transcripts in the aged human brain." J Neurochem. 59. 1117-1125 (1992)

[Publications] Yanagisawa K et al.: "Secretory pathway of β/A4 amyloid protein precursor in familial Alzheimer's disease with Val_<717> to Ile mutation." Neurosci Lett. 144. 43-45 (1992)

[Publications] Tsuji S et al.: "Molecular cloning ofhuman growth inhibitory factor cDNA and its down-regulation in Alzheimer's disease." EMBO J. 11. 4843-4850 (1992)

[Publications] Oyama F et al.: "β-Amyloid protein precursor and tau mRNA levels versus β-amyloid plaque and neurofibrillary tangles in the aged human brain." J Neurochem. 60. 1658-1664 (1993)

[Publications] Hasegawa M et al.: "Characterization of two distinct monoclonal antibodies to paired helical filaments(PHF)-Further evidence for fetal-type phosphorylation of the tau in PHF-." J Neurochem. 60. 2068-2077 (1993)

[Publications] Morishima M et al.: "Ubiquitin is conjugated with amino-terminally processed tau in paired helical filaments." Neuron. 10. 1151-1160 (1993)

[Publications] Yoshida H et al.: "tau in paired helical filaments(PHF)is functionally distinct from fetal tau.-assembly incompetence of PHF-tau-." J Neurochem. 61. 1183-1186 (1993)

[Publications] Kobayashi H et al.: "Molecular cloning of rat growth inhibitory factor cDNA and the expression in the central nervous system." Mol Brain Res. 19. 188-194 (1993)

[Publications] Watanabe A et al.: "In vivo phosphorylation sites in fetal and adult rat tau." J Biol Chem. 268. 25712-25717 (1993)

[Publications] Ueda K et al.: "Molecular cloning of a novel component of amyloid in Alzheimer's disease." Proc Natl Acad Sci USA. 90. 11282-11286 (1993)

[Publications] Oyama F et al: "Down's syndrome:Upregulation of β- amyloid protien precursor and tau mRNAs and their defective coordinatioin." J Neurochem. (in press).

井原康夫東京大学, 医学部(医), 教授 (60114386)