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The native state of prion protein (PrP) directly inhibits formation of PrP-amyloid fibrils in vitro

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Title

The native state of prion protein (PrP) directly inhibits formation

of PrP-amyloid fibrils in vitro( 内容と審査の要旨(Summary) )

Author(s)

本田, 諒

Report No.(Doctoral

Degree)

博士(医学) 甲第1059号

Issue Date

2018-03-25

Type

博士論文

Version

ETD

URL

http://hdl.handle.net/20.500.12099/75196

※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。

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 Ặྡ㸦ᮏ⡠㸧  Ꮫ ఩ ࡢ ✀ 㢮  Ꮫ఩ᤵ୚␒ྕ  Ꮫ఩ᤵ୚᪥௜  Ꮫ఩ᤵ୚せ௳  Ꮫ఩ㄽᩥ㢟┠  ᑂ ᰝ ጤ ဨ ᮏ ⏣ ㄹ㸦኱㜰ᗓ㸧 ༤ ኈ㸦་Ꮫ㸧 ⏥➨  ྕ ᖹᡂ  ᖺ  ᭶  ᪥ Ꮫ఩つ๎➨㸲᮲➨㸯㡯ヱᙜ

The native state of prion protein (PrP) directly inhibits formation of PrP-amyloid fibrils in vitro.

㸦୺ᰝ㸧ᩍᤵ ୰ᕝ ᩄᖾ 㸦๪ᰝ㸧ᩍᤵ ୗ⏿ ாⰋ  ᩍᤵ ୰ᓥ ⱱ ㄽ ᩥ ෆ ᐜ ࡢ せ ᪨  ࣄࢺ࡟࠾ࡅࡿࢡࣟ࢖ࢶࣇ࢙ࣝࢺ࣭ࣖࢥࣈ⑓ࡸ࢘ࢩ࡟࠾ࡅࡿ∵ᾏ⥥≧⬻⑕࡟௦⾲ࡉࢀࡿࡼ࠺࡟㸪ఏ㐩 ᛶᾏ⥥≧⬻⑕࡜ࡣ㸪⬻࡟ᾏ⥥≧⑓ኚࢆ♧ࡋ㸪࠿ࡘ௚ಶయ࡟ఏ᧛ྍ⬟࡞⑌ᝈ⩌ࢆᣦࡍࠋࡇࢀࡽࡢ⑌ᝈࡣ㸪 ⬻ෆ࡟Ꮡᅾࡍࡿṇᖖᆺࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ 3U3 ࡀ㸪␗ᖖᆺࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ 3U36F ࡟ㄗࡗ࡚ᢡࡾ ␚ࡲࢀࡿࡇ࡜࡟ࡼࡗ࡚⏕ࡌࡿࠋ3U3 ࡜ 3U36Fࡣ㸪඲ࡃྠࡌ࢔࣑ࣀ㓟㓄ิࢆ᭷ࡋ࡚࠸ࡿࡀ㸪ࢱࣥࣃࢡ㉁ࡢ ᢡࡾ␚ࡳ᪉ࡀ␗࡞ࡗ࡚࠾ࡾ㸪๓⪅࡟ࡣȘ࣊ࣜࢵࢡࢫᵓ㐀㸪ᚋ⪅࡟ࡣșࢩ࣮ࢺᵓ㐀ࡀ୺࡟ㄆࡵࡽࢀࡿࠋ 3U36Fࡣ㸪3U3 ࢆ 3U36F࡟ᵓ㐀ኚ᥮ࡍࡿస⏝㸦⮬ᕫ」〇స⏝㸧ࢆ᭷ࡋ࡚࠾ࡾ㸪ࡇࢀࡀఏ㐩ᛶᾏ⥥≧⬻⑕

ࡢឤᰁᛶࡢᇶ♏࡟࡞ࡗ࡚࠸ࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋ㏆ᖺ㸪ࡇࡢࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ឤᰁᶵᵓࡣ㸪௚ࡢ⚄ ⤒ኚᛶ⑌ᝈ̿̿౛࠼ࡤ࢔ࣝࢶࣁ࢖࣐࣮⑓㸪ࣃ࣮࢟ࣥࢯࣥ⑓࡞࡝̿̿ࡢ⑓ែࢆㄝ᫂ࡍࡿ௬ㄝ࡜ࡋ࡚ࡶὀ ┠ࢆᾎࡧ࡚ࡁ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ឤᰁᶵᵓࡢヲ⣽࡞࣓࢝ࢽࢬ࣒ࡢゎ᫂ࡣ㸪ఏ㐩 ᛶᾏ⥥≧⬻⑕ࡢࡳ࡞ࡽࡎ㸪⚄⤒ኚᛶ⑌ᝈ඲యࢆ⌮ゎࡍࡿୖ࡛㔜せ࡛࠶ࡿࠋ  ᮏ◊✲࡛ࡣ㸪ヨ㦂⟶ෆ࡟࠾࠸࡚㸪3U36Fࡀ࡝ࡢࡼ࠺࡟⮬ᕫ」〇ࡍࡿ࠿ࢆ㸪཯ᛂ㏿ᗘㄽⓗᡭἲࢆ⏝࠸࡚ ゎᯒࡋࡓࠋࡇࢀࡲ࡛㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇཯ᛂࡣᐇ㦂ື≀ࡸࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ឤᰁ⣽⬊ࢆ⏝࠸ ࡚ࡼࡃㄪ࡭ࡽࢀ࡚ࡁࡓࡀ㸪཯ᛂ㏿ᗘㄽⓗᡭἲࢆ⏝࠸ࡓᐃ㔞ⓗ࡞ゎᯒࡣ࡯࡜ࢇ࡝࡞ࡉࢀ࡚࠸࡞࠸ࠋᮏ◊ ✲࡛ࡣ㸪≉࡟㸪3U3 ࡢኳ↛≧ែࡀ㸪」〇཯ᛂ࡟࡝ࡢࡼ࠺࡟㛵୚ࡍࡿ࠿࡟㔜Ⅼࢆ⨨࠸࡚ゎᯒࢆ⾜ࡗࡓࠋ 3U3 ࡣ㸪⏕⌮ⓗ⎔ቃ࡟࠾࠸࡚ࡣ㸪ኳ↛≧ែ࡜ኚᛶ≧ែ㛫ࡢᖹ⾮ࡢୗ࡛Ꮡᅾࡋ࡚࠸ࡿࠋ⌧ᅾࡢ㏻ㄝⓗ࡞ ⪃࠼᪉࡛ࡣ㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇཯ᛂࡢᇶ㉁࡜ࡋ࡚ാࡃࡢࡣኚᛶ≧ែ࡛࠶ࡗ࡚㸪ࡇࢀ࡟ᑐࡋ࡚㸪 ኳ↛≧ែࡣ」〇཯ᛂࡢᇶ㉁࡜ࡣ࡞ࡾ࠼ࡎ㸪࠶ࡃࡲ࡛ኚᛶ≧ែࡢ๓㥑≧ែ࡜ࡋ࠿⪃࠼ࡽࢀ࡚࠸࡞࠸ࠋࡋ ࠿ࡋ㸪ࡑࡢ୍᪉࡛㸪ኚᛶ๣ࡸ S+ ࡞࡝࡛ኳ↛≧ែࢆ୙Ᏻᐃ໬ࡉࡏࡿ࡜㸪」〇཯ᛂࡀⴭࡋࡃຠ⋡໬ࡉࢀ ࡿࡇ࡜ࡶ♧ࡉࢀ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚㸪3U3 ࡢኳ↛≧ែࡣ㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇཯ᛂ࡟ᢚไⓗ࡟ാ ࠸࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡾ㸪ࡑࡢヲ⣽࡞࣓࢝ࢽࢬ࣒ࡢゎ᫂ࡀᮃࡲࢀࡿࠋ  ࠙ᑐ㇟࡜᪉ἲࠚ  ࡣࡌࡵ࡟ࣄࢺ 3U3 ࢆ㑇ఏᏊ⤌ࡳ᥮࠼኱⭠⳦⣔ࢆ⏝࠸࡚Ⓨ⌧ࡋ㸪࢔ࣇ࢕ࢽࢸ࢕࣮ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮ ࡸ㧗ᅽࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮࡞࡝ࢆ⏝࠸࡚㧗⣧ᗘࡲ࡛⢭〇ࡋࡓࠋࡇࡢࣜࢥࣥࣅࢼࣥࢺ 3U3㸦U3U3㸧࡟㉸ 㡢Ἴࢆ↷ᑕࡋ㸪3U36F࡟㢮ఝࡋࡓ⮬ᕫ」〇ᶵ⬟ࢆࡶࡘ࢔࣑ࣟ࢖ࢻ⥺⥔ࢆ⏕ᡂࡋࡓࠋḟ࡟㸪⏕ᡂࡋࡓ࢔࣑ ࣟ࢖ࢻ⥺⥔࡜ U3U3 ࢆΰྜࡋ㸪ࡇࡢ཯ᛂ⁐ᾮ୰࡛ࡢ࢔࣑ࣟ࢖ࢻ⥺⥔ࡢ」〇㏿ᗘࢆ㸪ࢳ࢜ࣇࣛࣅࣥ㹒ࡸ ࢤࣝℐ㐣ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮࡞࡝ࢆ⏝࠸࡚ㄪ࡭ࡓࠋࡉࡽ࡟㸪཯ᛂ⁐ᾮ୰ࡢ࢔࣑ࣟ࢖ࢻ⥺⥔⃰ᗘ㸪U3U3 ⃰ᗘ㸪࠾ࡼࡧࢢ࢔ࢽࢪࣥሷ㓟ሷ⃰ᗘࢆኚ໬ࡉࡏ㸪ࡇࢀࡽࡀ࢔࣑ࣟ࢖ࢻ」〇㏿ᗘ࡜࡝ࡢࡼ࠺࡟┦㛵ࡍࡿ ࠿ࡶㄪ࡭ࡓࠋ᭱ᚋ࡟㸪ᚓࡽࢀࡓᐇ㦂ࢹ࣮ࢱࢆ㓝⣲཯ᛂ㏿ᗘㄽࢆ⏝࠸࡚⌮ㄽⓗ࡟ゎᯒࡋ㸪ᐇ㦂⤖ᯝ࡟᭱ ࡶ㐺ྜࡍࡿࣔࢹࣝࢆᶍ⣴ࡋࡓࠋ

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࠙⤖ᯝࠚ  ࡣࡌࡵ࡟㸪ኳ↛≧ែኚᛶ≧ែ㛫ࡢᖹ⾮ࡀኚᛶ≧ែഃ࡟ᙉࡃഴ࠸ࡓ᮲௳ୗ㸦 PRO/ ௨ୖࡢࢢ࢔ ࢽࢪࣥሷ㓟ሷᏑᅾୗ㸧࡛࢔࣑ࣟ࢖ࢻ」〇㏿ᗘࢆ ᐃࡋࡓ࡜ࡇࢁ㸪࢔࣑ࣟ࢖ࢻ」〇㏿ᗘࡣῧຍࡋࡓ U3U3 ⃰ᗘ࡟ᑐࡋ࡚࣑࢚࢝ࣜࢫ࣓ࣥࢸࣥᆺࡢ㣬࿴᭤⥺ࢆ♧ࡋࡓࠋࡇࢀࡣᚑ᮶ᆺࡢ཯ᛂࣔࢹࣝ㸪ࡍ࡞ࢃࡕ 3U3 ࡢάᛶ୰㛫య 3U3 ࡀ࢔࣑ࣟ࢖ࢻ࡜」ྜయࢆᙧᡂࡋ㸪ḟ࠸࡛␗ᖖᆺ࡟ᵓ㐀ኚ᥮ࡉࢀࡿ࡜࠸࠺ࣔࢹࣝ࡜ ࡼࡃ୍⮴ࡋࡓࠋࡇࡢ⤖ᯝࡣ㸪ᑡ࡞ࡃ࡜ࡶ㸪ᚑ᮶ᆺࡢࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࣔࢹࣝࡀ㸪ᑡ࡞ࡃ࡜ࡶኳ ↛≧ែࡀࢃࡎ࠿ࡋ࠿Ꮡᅾࡋ࡞࠸ሙྜ࡟ࡣ㸪ṇࡋ࠸ࡇ࡜ࢆ♧ࡋ࡚࠸ࡿࠋ  ḟ࡟ኳ↛≧ែኚᛶ≧ែ㛫ࡢᖹ⾮ࡀኳ↛≧ែഃ࡟ഴ࠸ࡓ᮲௳ୗ㸦 PRO/ ௨ୗࡢࢢ࢔ࢽࢪࣥሷ㓟 ሷᏑᅾୗ㸧࡛ྠᵝࡢᐇ㦂ࢆ⾜ࡗࡓ࡜ࡇࢁ㸪ୖࡢ⤖ᯝ࡟཯ࡋ࡚㸪࢔࣑ࣟ࢖ࢻ」〇㏿ᗘࡣῧຍࡋࡓ 3U3 ⃰ ᗘ࡟ᑐࡋ࡚ᇶ㉁㜼ᐖᆺࡢ XSDQGGRZQ ᭤⥺ࢆ♧ࡋࡓࠋࡇࡢᇶ㉁㜼ᐖᆺ᭤⥺ࡣ㸪ኳ↛≧ែഃ࡟ᖹ⾮ࡀࡼ ࡾࢩࣇࢺࡍࡿ࡯࡝㸪ࡼࡾ㢧ⴭ࡜࡞ࡗࡓࠋࡇࡢ⤖ᯝࡣ㸪ᚑ᮶ᆺࡢ཯ᛂࣔࢹ࡛ࣝࡣㄝ᫂ᅔ㞴࡛࠶ࡾ㸪᪂ࡋ ࠸ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࣔࢹࣝࡢᏑᅾࢆ♧၀ࡍࡿࡶࡢ࡛࠶ࡗࡓࠋ㓝⣲཯ᛂ㏿ᗘㄽ࡟࠾ࡅࡿᇶ㉁㜼ᐖ ࣔࢹࣝ࠿ࡽࡢ㢮᥎࡟ࡼࡾ㸪ኳ↛≧ែ࡟ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࡢ୙➇ྜ㜼ᐖຠᯝࢆ௬ᐃࡋࡓࣔࢹࣝࢆ స〇ࡋࡓ࡜ࡇࢁ㸪ᐇ㦂⤖ᯝ࡜ࡼࡃ୍⮴ࡋࡓࠋࡲࡓ㸪ࡇࢀ࡜㢮⦕ࡍࡿࣔࢹࣝࢆ」ᩘಶ᳨ウࡋࡓ࡜ࡇࢁ㸪 ኳ↛≧ែ࡟ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࡢ㠀➇ྜ㜼ᐖຠᯝࢆ௬ᐃࡋࡓሙྜࡶྠᵝ࡟㸪ᐇ㦂⤖ᯝࢆࡼࡃㄝ᫂ ࡛ࡁࡿࡇ࡜ࡀࢃ࠿ࡗࡓࠋ  ࠙⪃ᐹࠚ  ᮏ◊✲ࡢ⤖ᯝࡣ㸪3U3 ࡢኳ↛≧ែ࡟㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࡢ㠀➇ྜཪࡣ୙➇ྜ㜼ᐖᶵ⬟ࡀ࠶ࡿ ࡇ࡜ࢆ♧၀ࡋ࡚࠸ࡿࠋࡇࢀࡣ㸪3U36F࡟஧ࡘࡢᶵ⬟ࡢ␗࡞ࡿ⤖ྜࢧ࢖ࢺ㸦άᛶ㒊఩࡜㜼ᐖ㒊఩㸧ࡀ࠶ࡿ ࡇ࡜ࢆព࿡ࡋ࡚࠾ࡾ㸪105 ࡸ⾲㠃ࣉࣛࢬࣔࣥඹ㬆ࢆ⏝࠸ࡓࡉࡽ࡞ࡿ◊✲ࡀᮃࡲࢀࡿࠋࡲࡓ㸪ᮏ◊✲ࡢ ⤖ᯝࡣ㸪3U3 ࡢኳ↛≧ែࢆ฼⏝ࡋ࡚㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ࡢ」〇ࢆຠ⋡ⓗ࡟ᢚไ࡛ࡁࡿྍ⬟ᛶࡶ♧ࡋ ࡚࠸ࡿࠋᐇ㝿࡟㸪⏕⌮ⓗ᮲௳ࢆ෌⌧ࡋࡓ᮲௳࡛ࢩ࣑࣮ࣗࣞࢩࣙࣥࢆ⾜ࡗࡓ࡜ࡇࢁ㸪㔝⏕ᆺࡢ 3U3 ࢆῧ ຍࡋࡓሙྜ㸪ᩘ wPRO/ ⛬ᗘࡢ⃰ᗘ࡛ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࢆᢚไ࡛ࡁࡿ࡜ࡢண ࢆᚓࡓࠋࡉࡽ࡟㸪 3U3 ࡢኳ↛≧ែࡢᏳᐃᛶࢆቑࡋࡓሙྜࢆࢩ࣑࣮ࣗࣞࢩࣙࣥࡋࡓ࡜ࡇࢁ㸪ࢃࡎ࠿ᩘ QPRO/ ⛬ᗘࡢ⃰ᗘ ࡛ࡶࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࢆຠ⋡ⓗ࡟ᢚไ࡛ࡁࡿ࡜ࡢண ࢆᚓࡓࠋ௨ୖ࠿ࡽ㸪ࢱࣥࣃࢡ㉁ᕤᏛࡸప ศᏊࣜ࢞ࣥࢻ➼ࢆ⏝࠸࡚ 3U3 ࡢኳ↛≧ែࢆᏳᐃ໬ࡉࡏ㸪ࡇࡢᏳᐃᆺ 3U3 ࢆ⏕య࡟ᢞ୚ࡍࡿࡇ࡜࡛㸪ࣉ ࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࢆຠ⋡ⓗ࡟ᢚไ࡛ࡁࡿྍ⬟ᛶࡀ♧ࡉࢀࡓࠋ  ࠙⤖ㄽࠚ  3U3 ࡢኳ↛≧ែࡣ㸪࢔࣑ࣟ࢖ࢻ」〇ࢆ㠀➇ྜࡲࡓࡣ୙➇ྜⓗ࡟㜼ᐖࡍࡿࠋ  ㄽ ᩥ ᑂ ᰝ ࡢ ⤖ ᯝ ࡢ せ ᪨  ⏦ㄳ⪅ ᮏ⏣ㄹࡣ㸪ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁ࡢ࢔࣑ࣟ࢖ࢻ」〇཯ᛂࢆ㏿ᗘㄽⓗᡭἲࢆ⏝࠸࡚ヲ⣽࡟ㄪ࡭㸪 ኳ↛≧ែࡢࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁࡟࢔࣑ࣟ࢖ࢻ」〇㜼ᐖຠᯝࡀ࠶ࡿࡇ࡜ࢆぢฟࡋࡓࠋᮏ◊✲ࡢᡂᯝࡣ㸪 ࣉࣜ࢜ࣥࢱࣥࣃࢡ㉁」〇ࡢ᪂ࡋ࠸཯ᛂࣔࢹࣝࢆ♧ࡍ࡜࡜ࡶ࡟㸪ఏ㐩ᛶᾏ⥥≧⬻⑕ࡢ᪂つ἞⒪ᡓ␎ࢆᥦ ᱌ࡍࡿࡶࡢ࡛࠶ࡾ㸪⚄⤒ᏛࡢⓎᒎ࡟ᑡ࡞࠿ࡽࡎᐤ୚ࡍࡿࡶࡢ࡜ㄆࡵࡿࠋ 㹙୺ㄽᩥබ⾲ㄅ㹛

Ryo P. Honda & Kazuo Kuwata: The native state of prion protein (PrP) directly inhibits formation of PrP-amyloid fibrils in vitro.

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