60
61
䛔䚹䛭䜜䛻ຍ䛘䠈
trans䉄 ring
ഃ䛷䛿ATP
䛾Ꮡᅾୗ䛷GroES
䛾⤖ྜ䛜㉳䛣䜛䛜䠈୍ᗘ⤖ྜ䛩䜛䛸䛭䜜௨㝆ゎ㞳䛫䛪䠈ඛ䛻ᇶ㉁䛜⤖ྜ䛧䛶䛔䛶䜒ᇶ㉁䛸ධ䜜௦䜟䜛䜘䛖䛻䛧䛶
GroES
䛜⤖ྜ䛩䜛䚹GroEL G192Xኚ␗య䛿
cis
䠈trans
䛹䛱䜙䛾䝸䞁䜾䛻䛚䛔䛶䜒GroES
䛿⤖ྜ䛷䛝 䜛䛜䠈䜻䝱䝡䝔䜱ෆ㒊䛻ᇶ㉁䛜ⴠ䛸䛧㎸䜎䜜䛺䛔䚹䛧䛛䛧䠈Gly192 ṧᇶ䜢⨨䛧䛯䜰䝭䝜㓟䛻䜘 䜚ᇶ㉁䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾άᛶ䛻㐪䛔䛜䜏䜙䜜䛯䚹69kDa 䛾⭠⳦⏤᮶䛾䝬䝹䝖䝕䜻䝇䝖䝸 䞁䜾䝹䝁䝅䝎䞊䝊(MalZ)䛿䠈GroEL䛾䜻䝱䝡䝔䜱ෆ㒊䛻ໟᣓ䛥䜜䛪䠈cis
䉄ringഃ䛻GroES
䛜䠈
trans
䉄ring
ഃ䛻MalZ
䛜⤖ྜ䛧䛯≧ែ䛷䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾䝰䝕䝹䛜ሗ࿌䛥䜜䛶䛔䜛(93)䚹GroEL G192X
ኚ␗య䛾cis
䉄ring 䛻䛿ATP
㠀Ꮡᅾୗ䛷GroES
䛜⤖ྜ䛩䜛䛾䛷MalZ
䛾䜘䛖䛻trans
䉄ring 䛻䛚䛔䛶GroES
䛜䜢䛧䛺䛔≧ែ䛷䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾ᶵᵓ䜒⪃䛘䜙䜜䜛䚹GroEL G192X ኚ␗య䛾䝅䝱䝨䝻䝙䞁ᶵ⬟䛾Ⓨ⌧䛻䛿
GroES
䛸䛾┦స⏝䛜䛝䛟㛵䛧䛶䛔䜛䛣䛸䛜⪃䛘䜙䜜䛯䛾䛷
QCM
ἲ䜢⏝䛧䛯ゎᯒ䜢⾜䛳䛯䚹㻌GroEL G192X
ኚ␗య䛿䠈ATP 䛾㠀Ꮡᅾୗ䛷䜒GroES
䛜⤖ྜ䛧䛶䛔䜛䛾䛜ぢ䜙䜜䛯(ᅗ 2-11)䚹䛥䜙䛻䛣䛾⤖ྜ㔞䛿⨨䛧䛯䜰䝭䝜㓟ṧᇶ䛜䛝䛔䜋䛹䜘䜚⤖ྜ䛧䛶䛔䜛䛾䛜᫂䜙䛛䛸䛺
䛳䛯䚹䛴䜎䜚䠈G192Xኚ␗య䛿ண䜑Open
≧ែ䛻䛺䛳䛶䛔䜛cis䉄 ring
ഃ䛻䛚䛔䛶ATP
䛾㠀 Ꮡᅾୗ䛷GroES
䛸┦స⏝䛩䜛䛣䛸䛜䛷䛝䠈Gly192 䜢䜘䜚䛝䛔䜰䝭䝜㓟ṧᇶ䛻⨨䛧䛯ኚ␗య䜋䛹
GroES
䛸┦స⏝䛧᫆䛔䛣䛸䛜☜ㄆ䛥䜜䛯䚹ᅗ2-12
䛻䛚䛔䛶GroEL WT
ཬ䜃GroEL G192N
ኚ␗య䛾ATP
Ꮡᅾୗ䠈㠀Ꮡᅾୗ䛻䛚䛡䜛GroES
䛸䛾┦స⏝ゎᯒ⤖ᯝ 䜢♧䛧䛯䚹䛣䛾⤖ᯝ䛻䛚䛔䛶䠈GroEL WT䛿ATP
㠀Ꮡᅾୗ䜘䜚䜒Ꮡᅾୗ䛾᪉䛜GroES
䛸⤖ྜ䛧䛶䛔䛯䚹䛣䜜䛿䠈GroEL WT 䛻
ATP
䛜⤖ྜ䛩䜛䛸Closed
≧ែ䛛䜙Open
≧ែ䛻䛺䜚GroES
䛜⤖ྜ䛷䛝䜛䜘䛖䛻䛺䜛ᛶ㉁䛜ᫎ䛥䜜䛶䛔䜛䛸⪃䛘䜙䜜䜛䚹GroEL G192Nኚ␗య䜒 㢮ఝ䛧䛯⤖ᯝ䜢♧䛧䛯䛣䛸䛛䜙䠈G192X ኚ␗య䛾trans䉄 ring
䛻䛚䛡䜛䜰䝢䜹䝹䝗䝯䜲䞁䛿ATP
㠀Ꮡᅾୗ䛷䛿Closed
≧ែ䛷䛒䜛䛜䠈ATP䛜⤖ྜ䛩䜛䛣䛸䛷Open
≧ែ䛸䛺䜚GroES
䛜⤖ྜ䛧䛯䛸᥎ 䛥䜜䜛䚹䛭䛾ᚋ䠈GroEL WT䛿ATP
䛜ຍỈศゎ䛥䜜ADP
䛻䛺䜛䛸GroES
䛜እ䜜䜛䚹䛧䛛䛧䠈G192X ኚ␗య䛿ᗘClosed
≧ែ䛻ᡠ䜛㝿䛻ᵓ㐀 ኚ 䛾ᨭⅬ䛸䛺䜛62
Hinge䊡㒊䛻䛚䛡䜛 Gly192
ṧᇶ䛾䜰䝭䝜㓟ṧᇶ䛜ኚ䛧䛯䛣䛸䛻䜘䜚ఱ䜙䛛䛾❧య㞀ᐖ䛜⏕䛨䜛䛣䛸䛷
Closed
≧ែ䛻ᡠ䜜䛪䠈GroES 䜒ゎ㞳䛧䛺䛟䛺䜛䛸⪃䛘䜙䜜䜛䚹GroEL 䜈䛾ኚ␗䜢ᑟධ䛩䜛䛣䛸䛷䠈ᮏ㉁ⓗ䛺ᵓ㐀䛜ኚ䛩䜛䛸䠈䝅䝱䝨䝻䝙䞁ᶵ⬟䛜䛝䛟ኚ䜟䜛䛣䛸䛻⧅䛜䜛 䛣䛸䛜⪃䛘䜙䜜䜛䚹䜎䛯䠈ATP 䛾ຍỈศゎ䛻䜘䜚
cis
䉄ring 䛾ᵓ㐀䛿ኚ䛧䛺䛔䛜䠈䝸䞁䜾㛫䛾 ᥋ゐ䛻䜘䜚trans
䉄ring 䛷䛾䝗䝯䜲䞁䛾ືస䛜⏕䛨䜛䛣䛸䛜䜽䝷䜲䜸㟁Ꮚ㢧ᚤ㙾䛻䜘䜚ゎᯒ䛥䜜 䛶䛚䜚䠈䛭䛾ᚋtrans
䉄ring 䛾ື䛝䛻䜘䜚య䛾䝸䞁䜾䛜ᒎ㛤䛧䠈ATP䛸ᇶ㉁䝍䞁䝟䜽㉁䛜⤖ྜ䛷䛝䜛䜘䛖䛻䛺䜛䛸♧䛥䜜䛶䛔䜛
(16)
䚹Open
≧ែ䛷䛒䜛cis
䉄ring༢⊂䛷䛾GroEL G192X
ኚ␗య䛾䝅䝱䝨䝻䝙䞁ᶵ⬟䜢䜘䜚ヲ⣽䛻ゎᯒ䛩䜛䛣䛸䜢┠ⓗ䛸䛧䠈GroEL䛾䝅䞁䜾䝹䝸䞁䜾ኚ␗య
SR-1 (89)䛻䛚䛡䜛 Gly192
ṧᇶ 䜢䛾䜰䝭䝜㓟ṧᇶ䛻⨨䛧䛯ኚ␗య䜢⏝䛔䛯 ᐃ䜢⾜䛳䛯䠄ᅗ2-13䠅䚹SR-1 G192X
ኚ␗య䛾
ATP
ຍỈศゎ⬟ຊ䛿GroEL WT
䛾༙ศ⛬ᗘ䛧䛛䛺䛔SR-1 WT
䜘䜚䜒䛥䜙䛻ప䛛䛳 䛯䚹䛧䛛䛧䠈GroES䛾ῧຍ䛷䜘䜚άᛶ䛜పୗ䛧䛶䛔䛯䛣䛸䛛䜙䠈GroES䛸䛾ぶᛶ䛜䛒䜛䛣䛸䛜♧䛥䜜䛯䚹Rhodanese 䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾άᛶ䛿
SR-1 WT
䜘䜚䜒ప䛔䛜Spontaneous
䜘 䜚䜒㧗䛔䛣䛸䛛䜙䜻䝱䝡䝔䜱ෆ㒊䜈ᇶ㉁䜢ྲྀ䜚㎸䜣䛷䛔䜛䛣䛸䛜⪃䛘䜙䜜䜛䚹䜎䛯䠈䝅䞁䜾䝹䝸䞁䜾 ኚ␗య䛿㏻ᖖ䝅䝱䝨䝻䝙䞁䝃䜲䜽䝹䛜୍ᗘ䛧䛛ᅇ䜙䛺䛔䛂䝅䞁䜾䝹䝍䞊䞁䜸䞊䝞䞊≧ែ䛃䛷䛒 䜛䛣䛸䛛䜙䠈䜻䝱䝡䝔䜱ෆ㒊䛻ⴠ䛸䛧㎸䜎䜜䛯ኚᛶMDH
䛿እ䛻ᨺฟ䛥䜜䛺䛟䛺䜛䛾䛷SR-1
ኚ␗య䛿
MDH
䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾ຓ䜢⾜䜟䛺䛔䛸䛥䜜䛶䛔䜛(94)䚹䛧䛛䛧䠈SR-1 G192Xኚ␗య䛾άᛶ䛿䝎䝤䝹䝸䞁䜾䛾
G192X
ኚ␗య䛾ሙྜ䛸䛿␗䛺䜚㧗䛔άᛶ䜢♧䛧䛶䛔䛯䚹䛣䜜䜘 䜚䠈SR-1 G192X ኚ␗య䛿ATP
䛾Ꮡᅾୗ䛻䛚䛔䛶䝅䞁䜾䝹䝍䞊䞁䜸䞊䝞䞊䛷䝃䜲䜽䝹䛜Ṇ䛫䛪䠈GroES䛜እ䜜䜛䛣䛸䛷
MDH
䛜እഃ䛻ᨺฟ䛥䜜䛶䛔䜛䛣䛸䜒⪃䛘䜙䜜䜛䚹䛭䛣䛷䠈ATP Ꮡᅾୗཪ䛿㠀Ꮡᅾୗ䛻䛚䛡䜛GroES
䛸䛾┦స⏝䛾ゎᯒ䜢⾜䛳䛯䚹ᅗ2-12
䛻䛚䛔䛶ATP
Ꮡᅾୗཪ䛿㠀Ꮡᅾୗ䛻䛚䛡䜛SR-1 G192I
䛸GroES
䛾┦స⏝ゎᯒ⤖ᯝ䜢♧䛧䛯䚹䛣䛾䜾 䝷䝣䜘䜚ྛ䜹䜲䝛䝔䜱䝑䜽䝟䝷䝯䞊䝍䞊䛾⟬ฟ䜢⾜䛳䛯(⾲2-2)䚹䛣䛾䝟䝷䝯䞊䝍䞊䛻䛚䛔䛶䠈SR-1 G2-2)䚹䛣䛾䝟䝷䝯䞊䝍䞊䛻䛚䛔䛶䠈SR-192I
ኚ␗య䛿ATP
䛾᭷↓䛻㛵䜟䜙䛪GroES
䛸䛾┦స⏝䛻ኚ䛿䜏䜙䜜䛺䛛䛳䛯䚹63
GroEL G192W
ኚ␗య䛾SPR
䛾⤖ᯝ䛸ẚ㍑䛧䛶SR-1 G192䊠ኚ␗య䛿ゎ㞳ᐃᩘ K d
䛜 ⴭ䛧䛟ᑠ䛥䛔䛣䛸䛛䜙GroES
䛜ゎ㞳䛧䜔䛩䛔䛣䛸䛜☜ㄆ䛷䛝䜛䚹SR-1 䛻䛚䛡䜛ᵝ䚻䛺㒊䛷 䛾Ⅼኚ␗䛿䛭䛾ศᏊ㔞䛻㐪䛔䛜⏕䛨䜛䚹䛣䜜䜙䛾ኚ␗య䛿ATP
䛾Ꮡᅾୗ䛷䛾GroES
䛾⤖ྜ䛜䛝䛟ῶᑡ䛧䛶䛔䜛䛜䠈mtMDH 䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾ຓ⬟ຊ䜢ಖᣢ䛧䛶䛔䜛䜒䛾䜒ሗ
࿌䛥䜜䛶䛔䜛
(95,96)
䚹䛣䛾䜘䛖䛻䠈SR-1 G192X
ኚ␗య䛿䠈SR-1 WT
䛸ẚ㍑䛧䠈Gly192
ṧᇶ 䜢ኚ䛥䛫䛯䛣䛸䛷GroES
䛸䛾┦స⏝䛜పୗ䛧䠈䛭䛾⤖ᯝMDH
䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾䜢⾜䛖䛣䛸䛜ྍ⬟䛻䛺䛳䛯䛸⪃䛘䜙䜜䜛䚹䛥䜙䛻䠈SR-1 䛻䛚䛡䜛
Gly192
䜢⨨䛧䛯䜰䝭䝜㓟ṧᇶ 䛻䜘䜚MDH
䛾䝸䝣䜷䞊䝹䝕䜱䞁䜾άᛶ䛻㐪䛔䛜䜏䜙䜜䛯䛣䛸䛛䜙䠈SR-1 G192X
ኚ␗య䛿GroES
䛸䛾┦స⏝䛾ᙉ䛥䛿Gly192
䜢⨨䛩䜛䜰䝭䝜㓟ṧᇶ䛻䜘䜚ኚ䛩䜛䛸⪃䛘䜙䜜䜛䚹 䛣䛾ᛶ㉁䛿䝎䝤䝹䝸䞁䜾䛾GroEL
䛻䛚䛡䜛cis
䉄ringഃ䛻㢮ఝ䛧䛯ᛶ㉁䛜ᫎ䛥䜜䛶䛔䜛䚹䛣 䛾SR-1 G192X
ኚ␗య䜢⏝䛔䛯ゎᯒ䛻䜘䜚䠈ATP 㠀Ꮡᅾୗ䛻䛚䛡䜛GroEL G192X
ኚ␗య䛾
cis -ring
䛻䛚䛔䛶䠈⨨䛧䛯䜰䝭䝜㓟ṧᇶ䛻䜘䛳䛶䛿䛺Open
≧ែ䜢䛸䛳䛶䛔䛺䛔 䛯䜑䛻GroES
䛸䛾┦స⏝䛜ᙅ䛔䛸⪃䛘䜙䜜䜛䚹䜎䛯䠈7 䛴䛾GroEL
䝃䝤䝴䝙䝑䝖䛻ᑐ䛧䛶GroES
䛾䝃䝤䝴䝙䝑䝖䛜䛶⤖ྜ䛧䛶䛚䜙䛪䠈GroEL-GroES䛾᥋ゐ㠃䛻䛚䛔䛶㝽㛫䛜⏕䛨䠈 䛭䛣䛛䜙ᇶ㉁䛜እ䛻ᨺฟ䛥䜜䜛䛣䛸䜒⪃䛘䜙䜜䜛䚹௨ୖ䛾䛣䛸䜘䜚䠈Gly192 䜢⨨䛩䜛䜰䝭䝜㓟ṧᇶ䜢䜘䜚㑅ᢥⓗ䛻䛩䜛䛣䛸䛷䜰䝢䜹䝹䝗䝯䜲䞁 䛾
Open
䛺≧ែ䛜ไᚚ䛥䜜䠈䛭䜜䛻క䛖GroES
䛸䛾┦స⏝䜔ኚᛶᇶ㉁䛾䝸䝣䜷䞊䝹䝕䜱䞁 䜾ຓ➼䛾䝅䝱䝨䝻䝙䞁䛸䛧䛶䛾ᶵ⬟䜢ㄪ⠇䛩䜛䛣䛸䛜ྍ⬟䛷䛒䜛䛣䛸䛜♧၀䛥䜜䛯䚹64
➨
➨ 3 ❶ GroEL Gly192 ኚ␗య䛻䜘䜛䜰䝭䝻䜲䝗⥺⥔จ㞟䛾
ᢚไຠᯝ
ドキュメント内
⺮ⓑ㉁ᕤᏛⓗᡭἲࢆ㥑ࡋࡓࢩࣕ࣌ࣟࢽࣥ GroEL ࡢ
(ページ 68-72)