94
95
䛯䜘䛖䛻䠈GroEL WT 䛿䠈䃐-Synuclein 䛾⥺⥔ᙧᡂ䛻䛚䛡䜛ึᮇ䛾ẁ㝵䛷ῧຍ䛩䜛䛣䛸䛷 α-Synuclein 䛜ᙧᡂ䛩䜛䜰䝭䝻䜲䝗⥺⥔ᮏయ䛾䛽䛨䜜䛯ᙧែ䛸䛿␗䛺䜚䠈䝥䝻䝖䝣䜱䝤䝸䝹䛜䛚
䛔䛻୪⾜䛻㓄⨨䛧䛯ᖹ䜙䛺ᙧែ䛷䛒䜛䛾䜢☜ㄆ䛧䛯䚹䛣䛾ᙧᡂ䛩䜛⥺⥔䛾ᙧែ䜢ㄪ⠇䛩䜛
⬟ຊ䛿䠈 α-Synuclein䛻ᑐ䛩䜛
GroEL WT
䛾᭱䜒ᙅ䛔┦స⏝䛜ᙳ㡪䛧䛶䛔䜛䛣䛸䛜⪃䛘 䜙䜜䜛䚹GroEL G192W
䛿α-Synuclein䛻ᑐ䛧䛶᭱䜒ᙉ䛔┦స⏝䜢♧䛧䛶䛔䛯䚹GroEL
䛸 䛾┦స⏝䛿䠈䃐-Synuclein 䛾༢㔞య䛻䛚䛔䛶ほ 䛧䛯䚹䃐-Synuclein 䛾䜰䝭䝻䜲䝗⥺⥔ᙧᡂᶵᵓ䛿䠈ጞ䜑䛻༢㔞యศᏊ䛜จ㞟䛧䠈䜸䝸䝂䝬䞊䛸䛺䜚䠈䛭䛣䛛䜙⥺⥔᰾䛜⏕䛨䠈䛣䛾⥺
⥔ ᰾ 䛻 ༢ 㔞 య ศ Ꮚ 䛜 ḟ 䚻 䛸 ⤖ ྜ 䛧 䛶䛔 䛟 䛣䛸 䛷 ⥺ ⥔ ఙ 㛗 䛜 ㉳ 䛣䜛 䛸 䛥䜜 䛶䛔 䜛 䚹䛴 䜎 䜚 䠈
Gly192
䜢 䛾 䜰 䝭 䝜 㓟 ṧ ᇶ 䛻 ⨨ 䛩 䜛 䛣䛸 䛷 䜘 䜚 䃐-Synuclein
䛸 䛾 ぶ ᛶ 䛜 ቑ ຍ 䛧 䛯GroEL G192X
ኚ␗య䛿䠈䃐-Synuclein ༢㔞యศᏊ䛸⤖ྜ䛩䜛䛣䛸䛷จ㞟䜢㜼ᐖ䛩䜛䛛䠈 ༢㔞య䛾⥺⥔᰾䜈䛾⤖ྜ䜢㜼ᐖ䛧䛶䛔䜛䛣䛸䛜⪃䛘䜙䜜䛯䚹䛥䜙䛻䠈ᅗ3-11
䛷Seed
Ꮡᅾୗ䛻䛚䛡䜛䃐-Synuclein䛾⥺⥔ᙧᡂ䛻䛚䛔䛶䠈GroEL WT 䛿
Seed
Ꮡᅾୗ䛷䃐-Synuclein 䛾⥺⥔ᙧᡂ䛜ຍ㏿䛥䜜䛯䚹䛧䛛䛧䠈GroEL G192W 䛿Seed
Ꮡᅾୗ䛻䛚䛔䛶䜒⥺⥔䛜ຍ㏿ 䛥䜜 䛺䛛䛳䛯 䚹䛣䛾 ⤖ ᯝ 䜘䜚䠈 ⥺ ⥔ ᰾ 䛻 ᑐ 䛩 䜛 䃐-Synuclein ༢ 㔞 య 䛾 ┦ స ⏝ 䜘䜚䜒
GroEL G192W
䛻ᑐ䛩䜛䃐-Synuclein༢㔞య䛾┦స⏝䛾᪉䛜䜘䜚ᙉᅛ䛷䛒䜛䛣䛸䛜⪃䛘 䜙䜜䛯䚹䛣䜜䜙䛾⤖ᯝ䜘䜚䠈GroEL 䛿䠈⥺⥔ᙧᡂ䛻䛚䛡䜛ึᮇ䛾ẁ㝵䛻ᙳ㡪䜢ཬ䜌䛧䠈GroEL 䛾
Hinge䊡㒊䛻䛚䛡䜛 Gly192
ṧᇶ䛻䛚䛡䜛䜰䝭䝜㓟ṧᇶ䜢⨨䛧䛯䛣䛸䛷䠈䜰䝢䜹䝹䝗䝯䜲䞁 䛾ഴ䛝䛸䃐-Synuclein ༢㔞య䜈䛾ぶᛶ䛻㐪䛔䛜⏕䛨䠈䛭䛾⤖ᯝ䃐-Synuclein 䛜ᙧᡂ䛩 䜛⥺⥔䛾ᙧែ䛻䜒ኚ䛜⏕䛨䛯䛣䛸䛜⪃䛘䜙䜜䜛䚹䛥䜙䛻䠈ᅗ3-8
䛷♧䛧䛯䜘䛖䛻䠈GroES 䛸A
䃑1-40䛻䛚䛡䜛GroEL G192W
䛾䜰䝭䝻䜲䝗⥺⥔ᙧᡂ䛾ᢚไ䛻䛴䛔䛶᳨ウ䛧䛯䛸䛣䜝䠈䛭䜜 䛮䜜䃐-Synuclein䛾ሙྜ䛸ྠᵝ䛻㧗䛔ᢚไຠᯝ䛜䜏䜙䜜䛯䚹A䃑1-40䛻䛚䛡䜛⥺⥔ᙧᡂ䛾Thioflavin-T
⺯ග䛿GroEL G192W
Ꮡᅾୗ䛷䛿GroEL WT
Ꮡᅾୗ䛸ẚ㍑䛧䛶⥺⥔᰾䛾 ᙧᡂ䛜㐜ᘏ䛧䠈䛥䜙䛻ᙧᡂ䛥䜜䜛⥺⥔㔞䜒ῶᑡ䛧䛶䛔䜛䜘䛖䛻䜏䜙䜜䛯䚹A䃑1-40䛿40
ṧᇶ䛾96
䜰䝭䝜㓟䛛䜙䛺䜛పศᏊ䝨䝥䝏䝗䛷䛒䜛䚹A䃑1-40 䛿
Closed
≧ែ䛾GroEL
䛷䜒䜻䝱䝡䝔䜱ෆ 㒊䛻ྲྀ䜚㎸䜎䜜䜛䛸⪃䛘䜙䜜䜛䛜䠈GroEL G192W ኚ␗య䛿䛥䜙䛻Open
≧ែ䛸䛺䜛䛣䛸䛷䜻 䝱䝡䝔䜱ෆ㒊䛻ྲྀ䜚㎸䜎䜜䜔䛩䛟䛺䜚䠈䜰䝭䝻䜲䝗⥺⥔ᙧᡂ䜢䜘䜚ᢚไ䛧䛶䛔䜛䛸⪃䛘䜙䜜䜛䚹䜎䛯䠈GroEL G192W
䛿ATP
䛾㠀Ꮡᅾୗ䛻䛚䛔䛶GroES
䛜⤖ྜ䛩䜛䛣䛸䜢௨๓ሗ࿌䛧䛶䛔䜛(19)
䚹GroES
䛿GroEL
䛾䜰䝢䜹䝹䝗䝯䜲䞁䛻䛚䛡䜛GroES
⤖ྜ㒊䛻⤖ྜ䛩䜛䚹GroES
䛾ᇶ㉁⤖ྜ㒊䛿ᇶ㉁⤖ྜ㒊䛻㏆䛔⨨䛻Ꮡᅾ䛧䛶䛔䜛(10)䚹䛥䜙䛻䠈GroES 䛾⥺⥔ᙧ ᡂ㐣⛬䛿䠈GroES䛜ኚᛶ䛩䜛䛣䛸䛷7
㔞యᵓ㐀䛜༢㔞య䛻ゎ㞳䛧䛯ᚋ䜰䝭䝻䜲䝗⥺⥔䜢ᙧᡂ 䛩䜛䜘䛖䛻䛺䜛(110)䚹ᅗ3-7
䛻䛚䛔䛶GroEL G192W
䛻GroES
䛜༢㔞య䛻ゎ㞳䛩䜛䜘䜚䜒᪩䛟⤖ྜ䛩䜛䛣䛸䛷䜰䝭䝻䜲䝗⥺⥔ᙧᡂ䜢ᢚไ䛧䛶䛔䜛ྍ⬟ᛶ䜒⪃䛘䜙䜜䜛䚹GroEL G192W 䛿䃐-Synuclein䠈GroES䠈A䃑1-40 䛻䛚䛡䜛䜰䝭䝻䜲䝗⥺⥔ᙧᡂ䜢ᢚไ䛧䛶䛔䜛䛣䛸䜢♧䛧䠈 ᵝ䚻䛺䜰䝭䝻䜲䝗⥺⥔䜢ᙧᡂ䛩䜛䝍䞁䝟䜽㉁䛾จ㞟ᢚไຠᯝ䜢᭷䛧䛶䛔䜛ྍ⬟ᛶ䛜⪃䛘䜙䜜䜛䚹 䛣䜜䜙䛾䜰䝭䝻䜲䝗⥺⥔䜢ᙧᡂ䛩䜛䝍䞁䝟䜽㉁䛻ᑐ䛧䛶
GroEL G192W
䛿Gly192
ṧᇶ䜢᭱䜒䛝䛔䜰䝭䝜㓟䛷䛒䜛
Trp
䛻⨨䛧䛯䛣䛸䛷ຠᯝⓗ䛺ᢚไ䜢♧䛧䛯䛣䛸䛛䜙䠈GroEL
䛾䜰䝢 䜹䝹䝗䝯䜲䞁䛿䜰䝭䝻䜲䝗⥺⥔ᢚไᶵᵓ䛻ᑐ䛧䛶㔜せ䛺ᙺ䜢ᯝ䛯䛧䛶䛔䜛䛸⪃䛘䜙䜜䜛䚹䛭䛾 䛯䜑䠈➨4
❶䛷䛿䃐-Synuclein䠈GroES䠈A䃑1-42䜢⏝䛔䛶䜰䝢䜹䝹䝗䝯䜲䞁༢⊂(GroEL-AD)䛻䜘䜛䜰䝭䝻䜲䝗⥺⥔ᙧᡂᢚไຠᯝ䛻䛴䛔䛶ゎᯒ䜢⾜䛖䚹
97
➨
➨ 4 ❶ 䝭䝙䝅䝱䝨䝻䞁䛂 GroEL-AD 䛃䜢⏝䛔䛯䜰䝭䝻䜲䝗⥺⥔
ᙧᡂᢚไຠᯝ
ドキュメント内
⺮ⓑ㉁ᕤᏛⓗᡭἲࢆ㥑ࡋࡓࢩࣕ࣌ࣟࢽࣥ GroEL ࡢ
(ページ 102-105)