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࣋ࣈࢲࢺᰶ㓗ࡡ᩺ᒈ㛜
̺ๆ⃥ᚺ➽ᛮᰶ㓗ࡡᵋ⠇̺
Functionalization of PNA
-A Novel Strategy for Active Control of DNA Recognition by External Factors using Peptide Ribonucleic
Acids (PRNA)-
ኬ㜨ኬᏕኬᏕ㝌ᕝᏕ◂✪⛁ฦᏄᏕᑍᨯ࣬57*894ࠔྙᠺ࡛โᚒࠕ/89࢙࣬ࣤࢹࣞࣅ࣭โᚒࣈࣞ .(475 /89
⏛ᙢ࣬୕
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9FNHKLNR<FIF࣬>RVKLKLVF.QRXH
ࠚ ྻ⏛ᕰᒜ⏛ୢ㸧̾㸦
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SFSHU\H\RXOIOLNHWRJL[HFQR[HU[LH\RIWKHVHVWUFWHJLHVFQISURSHUWLHVRI53&IHUL[FWL[HV
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2
ࡢࡋࡴ࡞
)3& 㔔ࡼࡎࢆᵋ㏸࣠ࢹࢮ࡛ࣤࢠࣛࢴࢠ࡞ࡻࡽᥞ᱄ࡈࡿ࡙࠾ࡼ ࿔ᖳ࡞࠵ࡒࡾ ᖳ ᭮ ࠉ
㐅ᅗᨳᗋࡼࡦ࡞⭶ኬᩐ࡞ࡡࡾ◂✪⩽ࡡ༝ງࡡᠺᯕ࡛ࡊ࡙ࣃࢹࢣࢿ࣑㒼าỬᏽゝ⏤ࡡᏰᐃ
ゕࡈࡿࡒࠊࡆࡡࢣࢿ࣑㒼าሒ࡞ᇱࡘࡀࠉᰶ㓗Ꮥ࣬࣋ࣈࢲࢺᏕ࣬⢶㙈ᕝᏕࢅ୯ᚨ࡛ࡊࡒ⏍Ꮥࠉ
⏍᭯ᶭᏕࠉ⏍᭯ᶭྙᠺᏕࠉ⏍మᮞᩩ⛁Ꮥࡢࠉࣆࣤࢠࢨࣘࢻࣜࢣࢿ࣑ࠉࣈࣞࢷ࢛࣭࣑ࠉࢡࣚ
ࢤ࣭࣑࡛⥪⛘ࡈࡿࡾࢣࢿ࣑ࢨ࣭ࢠ࢙ࣤࢪࢸ࣭ࢰ࡞ᇱࡘ࠷ࡒ᩺ࡊ࠷◂✪࡛ࡊ࡙ࡻࡽୌᒒἸ┘ࡈࡿࠉ᭩
ࡵ⢥ງⓏ࡞◂✪ࡈࡿࡾฦ㔕ࡡୌࡗ࡛ࡖ࡙࠷ࡾࠊࡆࡡࡻ࠹⫴ᬊࡡࡵ࡛ࠉᮇ✇࡚ࡢࣆࣤࢠࢨࣘࢻࣜࢣ
ࢿ࣑࣬ᶭ⬗ᛮᰶ㓗ᵋ⠇ࢅ┘ᣞࡊࡒᰶ㓗Ꮥࠉ≁࡞ྙᠺᏕⓏふⅤ࠾ࡼᰶ㓗ᵋ㏸ࢅಞ㣥࣬ᨭንࡊࡒࠔெᕝ
ᰶ㓗ࠕ࡛⥪⛘ࡈࡿࡾᰶ㓗࣓ࢸࣜ࡞㛭ࡌࡾ◂✪ࡡୌ㒂ࢅ⣺ࡊࡒ࠷ࠊ࠽⌟ᅹ᭩ࡵ᭯ྞࠔெᕝᰶ㓗ࠕ࡛
ࡊ࡙▩ࡼࡿ࡙࠷ࡾ࣋ࣈࢲࢺᰶ㓗㸝53&㸞࡞㛭ࡊ࡙ࡢࠉ
ࠔ᭯ᶭྙᠺᏕ༝ఌヽࠕ ᖳ ᭮ྒ࡞ᮟ୕࣬ᒷ℡
ࡼ࡞ࡻࡽレࡊ࠷⥪ྙㄵᩝሒ࿈ࡈࡿ࡙࠷ࡾࡡ࡚ ࠉᮇ✇࡚ࡢ᭩ิ࡞ࠔெᕝᰶ㓗ࠕ࡞㛭ࡌࡾ◂✪ࢅᴣㄕࡊࠉ
53& ࡞㛭ࡊ࡙ࡢ ᖳ௧㜾ࡡࢸ࣭ࢰ࣭ࢅ୯ᚨ࡞⣺ࡊࡒ࠷ࠊ᭩ᚃ࡞አ㒂ᅄᏄ࣬አ㒂ๆ⃥ᚺ➽ᛮெᕝᰶ㓗
ᵋ⠇ࢅ┘ᣞࡊࡒᠻࠍࡡ◂✪ࢅ⣺ࡌࡾࠊ
ᫎᖳࣃࢹධࢣࢿ࣑㒼าỬᏽᏰࡊࠉࡐࡡよᯊ㐅ࡳࡆ࡛࡞ࡻࡽ㟸ᖏ࡞ኣࡂࡡ㔔さሒᚋࡼࡿࡾ
ࡆ࡛ᙽࡈࡿ࡙࠷ࡾࠊࡌ࡚࡞ᬳᏽࢣࢿ࣑ࡡよᯊ⤎ᯕ࡞ࡻࡽ ࠉኣࡂࡡ⑄ᝀ࣬⑄㐿ఎᏄࠉ࢝ࣤ㐿ఎᏄ
ࡡᅄ㐿ఎᏄࡡ㒼า᪺ࡼ࠾࡛ࡈࡿࠉ⑄࣬⑄ᝀࡡⓆ࡛ᑊ㇗㐿ఎᏄࡡୌሲᇱኣᆵ㸝835V㸞ࢅ୯ᚨ࡛
ࡊࡒ㐿ఎᏄ㒼าኣᆵ࡛ࡡ┞㛭⌟ᅹ⢥ງⓏ࡞◂✪ࡈࡿ࡙࠷ࡾࠊᅄ㐿ఎᏄ⩄ࡡ 835V ࡛Ⓠ࡛ࡡ┞㛭᪺
ࡼ࠾࡞ࡿࡣࠉ㐿ఎᏄチ᩷ࡷⷾវུᛮ࡞ᑊࡌࡾῼ࣬ึืྊ⬗࡛ࡽࠉ࢛࣭ࢱ࣭࣒ࢺ༈⒢࣬㜭
⒢ࡡᐁ⌟ᙽࡈࡿࡾࠊࡈࡼ࡞ࡆࡿࡼࡡ㐿ఎሒ࡞ᇱࡘࡀࠉḗୠࡡ⒢Ἢ࡛ࡊ࡙Ἰ┘ࡈࡿ࡙࠷ࡾ✒
ᴗⓏ㐿ఎሒⓆ⌟โᚒ࡞ᇱࡘࡂ㐿ఎᏄ⒢Ἢࡡᐁ⌟ࡵᙽࡈࡿࡾࠊ㐿ఎᏄ⒢࡞ࡢࢸࢼࣤࢸ࣐ࢻ
࣭ࢭḖࡡᝀ⩽࡞ᑊࡌ
ࡾ⒢࡚ᗀࡂ▩ࡼࡿࡾࡻ
Modified Nucleotide
NH2
࠹࡞ࡖࡒ㐿ఎᏄᑙථἪ
N
ࡡ࡞ࠉአ㒂࠾ࡼຊ࠻ࡒ
ሲᇱ㒼า≁␏Ⓩᰶ㓗⤎ྙ
ฦᏄ࡞ࡻࡽ⑄࣬⑄ᝀ≁
᭯ࢰࣤࣂࢠ㈻ྙᠺࡷ࢝ࣤ
⣵⬂ࡡቌᖕ࡞ྊḖ
O
O
O P S - O P CH3
O
O
ࢰࣤࣂࢠ㈻ྙᠺࢅ
P73& ࡡṹ㝭࡚ᢒโࡌࡾ
ࡆ࡛࡞ࡻࡽ⒢ࡌࡾࠉ
ࣤࢲࢬࣤࢪ 73& Ἢࡷࠉࡻ
N
N
O
O P OO
O
B
O
ᰶ㓗ㄇᑙమ࡞⾪ࡈࡿࡾ
O
㝭ࢅ㜴ᐐࡌࡾࡆ࡛࡞ࡻࡽ
modified bases
O
O
O P OO
O R
O
O P N + S P SO R
O
etc.
ࡽ᰷ᮇⓏ࡞ᰶහࡡ㔔ࡼ
ࡎࢆ )3& ࡞⤎ྙࡊ㌹ṹ
N
O
B
O
O
B
OMe or Et
B
O
O P O-
etc.
Chart 1 Structure of modified nucleotides.
⒢ࡊࡻ࠹࡛ࡌࡾࣤࢲࢩѸࣤᰶ㓗Ἢ᳠ゞࡈࡿ࡙࠷ࡾࠊࣤࢲࢩѸࣤᰶ㓗ἪࡢࠉኬࡀࡂࣤࢲࢬࣤࢪἪ
3
࡞ྱࡱࡿࡾࡆ࡛ࡵኣ࠷ࠊ ᖳᚃ༖࡞࢛ࣛࢥࢽࢠ࢛ࣝࢲࢺࡷಞ㣥࢛ࣛࢥࢽࢠ࢛ࣝࢲࢺࢅ⏕࠷ࡒࣤࢲ
ࢬࣤࢪἪࡡ᳠ゞ࡞ࡻࡽࠉࡐࡡ᭯⏕ᛮୌ㒂ᐁッࡈࡿࡒࠉ㓕⣪ฦよ࡞ࡻࡽࣤࢲࢬࣤࢪฦᏄ࡛ࡊ࡙
⏕ࡈࡿࡒ࢛ࣛࢥࢽࢠ࢛ࣝࢲࢺ㢦㎷㏷࡞ฦよࡈࡿ࡙ࡊࡱ࠹ࡒࡴ්⌟ᛮ࡞இࡊࡂࠉᏭᏽࡊࡒᶭ⬗Ⓠ⌟ࡢ
ሒ࿈ࡈࡿ࡙࠷࠷ࠊࡆࡿࡼࣤࢲࢬࣤࢪἪࡢᰶ㓗࡚ࣝ࣊ࣜࡡࢰࣤࣂࢠ㈻ྙᠺᢒโࢅ┘Ⓩ࡛ࡊ࡙࠷ࡾࡒࡴࠉ
㐿ఎᏄࡡ々⿿࣬㌹࣬⩳ゼࡡࡡ㐛⛤ࢅ㜴ᐐࡌࡾࡆ࡛ࡵྊ⬗࡚ࠉࢣࢿ࣑ࢨ࣭ࢠ࢙ࣤࢪ࡞ᇱࡘࡂ༈Ꮥฦ㔕
ࡡᚺ⏕ࡢࡵࡔࢀࢆࠉฦᏄ⏍∸Ꮥࡷ㐿ఎᏄᕝᏕᵕࠍฦ㔕࠾ࡼἸ┘ࡈࡿ࡙࠷ࡾࠊࣤࢲࢬࣤࢪฦᏄ
࡛ࡊ࡙࢛ࣛࢥࢽࢠ࢛ࣝࢲࢺࢅ⏕࠷ࡒ㝷ࡡ᰷ᮇⓏၡ㢗ࢅよỬࡌࡾࡒࡴࠉ㸦㸞㧏࠷ᰶ㓗ሲᇱ㒼าヾㆉ⬗ࠉ
㸧㸞ࢰ࣭ࢣࢴࢹ )3&࣬73& ࡛ࡡࢤࣤࣈࣝࢴࢠࢪࡡ㧏࠷Ꮽᏽᛮࠉ㸨㸞ᏕⓏࠉ≁࡞㓕⣪Ⓩ㸝⣵⬂හ㸞Ꮽᏽ
ᛮࠉ㸩㸞73& ࡛ࡡࢤࣤࣈࣝࢴࢠࢪ 73FVH- ࡡᇱ㈻࡛ࡾࠉ㸪㸞㧏࠷⣵⬂⭯㏩㐛ᛮࠉ㸫㸞ࢰࣤࣂࢠ㈻
Ꮛᅹୖ࡞࠽ࡄࡾᰶ㓗࡛ࡡ㐽ᢝⓏ┞பష⏕ᛮࠉࡡ≁ᛮࢅ᭯ࡌࡾࣤࢲࢬࣤࢪฦᏄࡡ⿿ࢅ┘ᣞࡊࠉኮ
↓ᰶ㓗ࡷ⮤ິྙᠺჹ࡞ࡻࡽᚋࡼࡿࡾࠔྙᠺᰶ㓗ࠕࡡሲᇱ㒂ࠉ⢶㒂ࠉࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㒂ࢅᏕಞ㣥ࡊࡒ
ࠔಞ㣥ᰶ㓗ࠕ࡛⥪⛘ࡈࡿࡾኣᩐࡡᰶ㓗ㄇᑙమࡷ㸝ࢲ࣭ࣔࢹ㸦㸞ࠉᰶ㓗ࡡࣆࣚࢿ࣭ࢪ̺ࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㦭
ࢅኬࡀࡂንᥦࡊࡒࠔெᕝᰶ㓗ࠕ
B
O
O
O
O
–O
୕ࡾྙ∸⩄ሒ࿈ࡈࡿ࡙࠷ࡾ
N
P O
B
PNA
H N
⣪ᶭ⬗ࢅ᭯ࡌࡾࣛ࣍ࢧ࣑ࡷ
H 2N
B
H2N
B
B
O
18)
18)
)
)
H N
H N
N
O
⏕ᒈ㛜ࡌࡾࡒࡴ࡞ࡢࠉ73&࣬)3&
ࡡ࣭ࣛ࣍ࢪ࣬ࢸ࢛࢞ࢨ࣭ࣛ࣍ࢪ
O
*
N
N
B
B
O
O
(
NH
O
B
)
)
N
H
O
N
O
ࡾ⁈よᛮࡂ᭯ᶭ⁈፳ࡡ῟ຊ
࡞ࡻࡽᐖ᪾࡞ንᛮࢅུࡄ࡙
B
B
O
aa1
aa2
O
aa3
)
O
O
H
)
22)
H N
HN
B
N
Ꮽᏽ࡚ࠉỀ௧አࡡ⁈፳࡞ᑊࡌ
)
21)
20)
)
19)
)
O
̺ࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㦭᭯ࡌ
ࡾࠉ㓕⣪⪇ᛮࡂᏕⓏ࡞ࡵ
O
(
)
)
O
B
N
O
࡙ࡀࡒࠊ࡛ࡆࢀᰶ㓗ࢅ⌟ᐁⓏ
ᶭ⬗ฦᏄ࣬ᶭ⬗ᮞᩩ࡛ࡊ࡙ᚺ
H N
B
*
ฦᏄࡵኣᩐ▩ࡼࡿࡾࡻ࠹࡞ࡖ
N
)
17)
)
NH
73&Lࠉࣈࢰ࣏࣭ࢅࡢࡋࡴ࡛ࡌ
ࡾኣᙤ࡚ࠉິⓏᶭ⬗ࢅ᭯ࡌࡾ
O
N
)
)
O
NH
O
H 2N
N
16)
)
H N
O
ࠊ
ୌ᪁ࠉᰶ㓗ࠉ≁࡞ 73& ࡞ࡢ㓕
5)
9)
)
)
RNA
O
)
DNA
⢥ງⓏ࡞᳠ゞࡈࡿࠉ⭶ኬᩐ࡞
O
O OH
P O
ࡡᵋ⠇ྙᠺᏕⓏぜᆀ࠾ࡼ
H N
)
–O
࡛⥪⛘ࡈࡿࡾᰶ㓗࣓ࢸࣜฦᏄ⣌
B
)
O
ࡊࡱ࠹࠷ࡂࡗࡵࡡၡ㢗ኬ
ࡀ㝸ᐐ࡛ࡾࠊࡱࡒ㐿ఎᏄ
O
⒢ⷾࡷ༈⏕ᮞᩩ࡞࠽࠷࡙ࡵࠉ⏍
)
మහᏭᏽᛮࡡḖዯኬࡀၡ
㢗ࢅᢢ࠻࡙࠽ࡽࠉ༎ฦᶭ⬗ࢅ
Chart 2 DNA, PNA, and PNA derivatives.
4
Ⓠ⌟ࡌࡾᐁ⏕Ⓩᮞᩩ㛜Ⓠ࡞ࡢ⮫ࡖ࡙ࡢ࠷࠷ ࠊ
ࡆࡿࡼࡡၡ㢗ࢅよỬࡊࠉᰶ㓗ࢅࡻࡽᏭᏽᶭ⬗ฦᏄ࡛ࡊ࡙Ὡ⏕ࡌࡾࡒࡴ࡞ࡢᰶ㓗ࡡᣚࡗ㦭ᵋ㏸ࢅࠉ
࢙ࢪࢷ࣐ࣜ࣬ࢺࡐࡊ࡙⅛⣪̺⅛⣪⤎ྙࡡࡻࡽᏭᏽ㦭ᵋ㏸࡞ንᥦࡊࡒㄇᑙమࡡᰶ㓗࣓ࢸࣜྙ
∸ࡡ㛜Ⓠᚪさ࡛ࡾࠊࡆࡡࡻ࠹ᰶ㓗࣓ࢸࣜࡢࠉࡡࣤࢲࢬࣤࢪฦᏄ࡛ࡊ࡙ࡵ㔔さ࡚ࠔெᕝᰶ㓗ࠕ
࡛⥪⛘ࡈࡿࡾࠊୌ᪁ࠉ࡞ࡵ㏑ࡒࣤࢲࢬࣤࢪฦᏄࠉࣤࢲࢩѸࣤࠉࢸࢤ 73& ࡛ࡊ࡙⌟ᅹᗀࡂ
⏕࠷ࡼࡿ࡙࠷ࡾࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㒂ࡡ㓗⣪ࢅ࢛ཋᏄࠉ࣐ࢿㄇᑙమ࡚⨠ᥦࡊࡒ SKRVSKRURWKLRFWH ࡷ
SKRVSKRUFPLIFWH ࡢࠉ㦭ᵋ㏸ࢅᏕንᥦࡊࡒࠔெᕝᰶ㓗ࠕ࡛༇ืࡌࡾࡒࡴࠔಞ㣥ᰶ㓗ࠕ࡛ࡣࡿࡾࡆ࡛
ኣ࠷ࠊ
ெᕝᰶ㓗ࡼࡦ࡞⛸ࠍࡡ 53& ㄇᑙమ
ெᕝᰶ㓗ࡢ )3& ࡡ㔔ࡼࡎࢆᵋ㏸Ⓠぜ࠾ࡼ⣑ ᖳᚃࡡ ᖳࡡ࠽ࢂࡽ࡞ /RQHV ࡛ <FONHU ࡼࠉ0F^H
ࡼࡐࡿࡑࡿࢽࢠ࢛ࣝࢨࢺࢅ⏕࠷ࡒ࣎ࣛࢽࢠ࢛ࣝࢲࢺ࣓ࢸࣜࢅሒ࿈ࡊࡒࡆ࡛࡞ࡢࡋࡱࡽࠉࡐࡡᚃ➁ᮇࠉ
4[HUGHUJHU5LWKF ࡼࡡࢡ࣭ࣜࣈᰶ㓗ሲᇱࢅᑙථࡊࡒࣄࢼ࣏ࣜ࣎ࣛѸࡷࠉ࣐࣎ࣛࢿ㓗ࢅ୯ᚨ࡞⛸ࠍࡡ
ᵋ㏸ࢅ᭯ࡌࡾㄇᑙమ㛜Ⓠࡈࡿࠉᶭ⬗ᛮ㧏ฦᏄ࡛ࡊ࡙ᕝᴏᮞᩩࢅ୯ᚨ࡞ᵕࠍᒈ㛜ሒ࿈ࡈࡿ࡙ࡀࡒ ࠊ
ࡊ࠾ࡊࠉெᕝᰶ㓗ࢅ⏕࠷ࡒ⏍మᶭ⬗㛭㏻Ꮥࡷ⏍Ꮥ࡞㛭ࡌࡾฦᏄ࡚ࣝ࣊ࣜࡡ◂✪ࡢ⢥ງⓏ࡞⾔ࢂࡿ
࡙࠷ࡾ࡛ࡢゕ࠷㞬࠷≟ឺ࡚࠵ࡖࡒࠊ
ࡊ࠾ࡊࠉ ᖳ࡞ 3LHOVHQ ࡼ࡞ࡻࡽ 3࣐ࢿ࢙ࢲࣜࢡࣛࢨࣤࢅ㙈㦭࡛ࡌࡾ࣋ࣈࢲࢺᰶ㓗
㸝5HSWLIH3XHOHLH&HLI 53&㸞ሒ࿈ࡈࡿ㸝ࢲ࣭ࣔࢹ 㸞ࠉ≟Ἓࡢୌንࡊࡒࠊ53& ࡢ┞Ⓩୌᮇ㙈 )3&
ࡷ 73&ࠉ≁࡞ 73& ࡛ <FWVRQ(ULHN ሲᇱᑊᙟᠺ࡞ᇱࡘࡂᏭᏽ㔔ࡼࡎࢆ㘊మࢅᙟᠺࡌࡾ࡛භ࡞ࠉࡼࡎࢆ
≟ࡡ㔔㙈㘊మࢅᙟᠺࡊ࡙࠷ࡾ )3& ࡛ࡵሲᇱ≁␏Ⓩ࡞┞பష⏕ࡊࠉ53&࣬)3& ᆵ㔔㙈㘊మࢅᙟᠺࡊࠉ)3&
㙈ࡢୌᮇ㙈≟ឺ࡛ࡾ VWUFQIILVSOFHHPHQW ሒ࿈ࡈࡿࡒࠊࡈࡼ࡞㒼า࡞ᑊᚺࡊ࡙ᵕࠍࢰࣈࡡ㔔
㙈㘊మࡡᙟᠺࡵሒ࿈ࡈࡿ࡙࠷ࡾ ࠊ53& ࡡᛮ㈻࡞ࡗ࠷࡙ࡡレ⣵よㄕࡢࡡ⥪ྙㄵᩝ ࡞㆙ࡾࠉ53& ࡡ
⾪Ⓩඁࡿࡒ≁ᛮࡢࠉ㸦㸞࣐ࢺ㦭ࢅ᭯ࡌࡾࡒࡴࢽࢠ࣭ࣝࢭ࡞ᑊࡊ࡙ࡢࡵࡔࢀࢆࠉࣈࣞࢷ࣭ࢭ
࡞ᑊࡊ࡙ࡵ⪇ᛮࢅ♟ࡊࠉ㧏࠷㓕⣪⪇ᛮࡼࡦ࡞ᏕⓏᏭᏽᛮࠉ㸧㸞53&࣬)3&ࠉ53&࣬73& 㘊మභ࡞ኮ
↓ᰶ㓗㘊మ࡞Ẓ㍉ࡊ࡙㟸ᖏ࡞㧏࠷Ꮽᏽᛮࠉ㸨㸞࣐ࢪ࣏ࢴࢲሲᇱᑙථ࡞ࡻࡽኬࡀࡂୖࡌࡾ㘊మᏭᏽᛮࠉ
ࡌࢂࡔ㧏࠷ሲᇱ㒼า㐽ᢝᛮࠉ࡚࠵ࡾࠊ㸦㸞53& ࡡᏭᏽᛮࡢฦᏄスゝ୕ࠉெᕝᰶ㓗࡛ࡊ࡙ࡢᙔ↓࡛ࡵ⩻࠻
ࡼࡿࡾࠉ㸧㸞53& )3& ࡛ᙟᠺࡌࡾ㘊మࡡ⼝よῺᗐ㸝9P㸞ࡢࠉᑊᚺࡌࡾ )3&࣬)3& 㘊మࡡࡐࡿ࡞Ẓ㍉ࡊ
࡙ୌሲᇱᑊ࠵ࡒࡽ ࠤΥࡵ㧏ࡂ㟸ᖏ࡞Ꮽᏽ࡚࠵ࡽࠉࡆࡿࡢ 53& ࢿࢼ࢛ࢼࢴࢠ࣐ࢺ㙈㦭ࢅ᭯ࡌ
ࡾࡒࡴࠉኮ↓ᰶ㓗㘊మࡡࡻ࠹࡞ࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㦭ࢼ࢛ࣤ㛣࡚ࡡ㟴㞹ⓏཬⓆᏋᅹࡊ࠷ࡒࡴ࡛⩻
࠻ࡼࡿࡾࠊࡆࡡࡆ࡛ࡢ㧏ሲ⃨ᗐ᮪௲ୖ࡚ኮ↓ᰶ㓗㘊మࡡᏭᏽᛮቌຊࡌࡾࡡ࡞ᑊࡊ 53&࣬)3&73& 㘊మࡡ
Ꮽᏽᛮࡢୖࡌࡾࡆ࡛࠾ࡼࡵ☔ヾ࡚ࡀࡾ ࠊࡈࡼ࡞ࠉ㸨㸞53&࣬)3& 㘊మ࡚ࡢ ࣧᡜࡡ࣐ࢪ࣏ࢴࢲሲᇱࡡ
Ꮛᅹ࡞ࡻࡽ 9P ࡢ ࠤΥࡵୖࡊࠉඁࡿࡒሲᇱ㒼าヾㆉᛮࢅ♟ࡌࡆ࡛ሒ࿈ࡈࡿ࡙࠷ࡾࠉࡆࡿࡢ 53&
ࡡ㙈㦭࣐࣎ࣛࢺ㦭࡚࠵ࡽẒ㍉Ⓩ๙├ᵋ㏸ࢅ᭯ࡊࠉᑻᡜⓏ㙈ࢤ࣒ࣤ࣌Ѹࢨࣘࣤን࣐ࢪ
࣏ࢴࢲሲᇱ࡞ࡻࡽㄇ㉫ࡈࡿࠉࡐࡡ⤎ᯕ࣐ࢪ࣏ࢴࢲሲᇱ㎾എࡡሲᇱᑊࡵよ㞫ࡌࡾࡆ࡛࡞ᇱࡘࡂ࡛⩻࠻ࡼࡿ
࡙࠷ࡾࠊ
ࡆࡡࡻ࠹࡞ඁࡿࡒ≁ᛮ࣬ᶭ⬗ࢅ♟ࡌ 53& ࡚࠵ࡾࠉ࣐࣎ࣛࢺ㙈࡞々⣪⎌࡚࠵ࡾᰶ㓗ሲᇱࢅࢡࣚࣆ
ࢹࡊࡒ࢛ࣛࢥ࣏࣭࡚࠵ࡾ࡛⩻࠻ࡾ࡛ᙔ↓࡚ࡢ࠵ࡾࠉỀ࡞ᑊࡌࡾ⁈よᛮẒ㍉Ⓩࡂࠉ⌟ᐁ࡞⏕࠷ࡼࡿ
ࡾࡡࡢ 㔖మ⛤ᗐࡱ࡚࡚ࠉ⮤ᕤఌྙᛮࡵᙁࡂỀᛮ࡞ᇱࡘࡂࢰࣤࣂࢠ㈻ࡷ㓕⣪㢦࡛ࡡ㟸≁␏Ⓩྺ╌ࡵ↋
5
っ࡚ࡀࡍࠉỀᛮ࡞㉫ᅄࡌࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾ⭯㏩㐛ᛮࡡḖዯ็ၡ㢗ࡵెࡎᣚࡗࠊࡱࡒࢰ࣭ࢣ
ࢴࢹ )3&73& ࡛ࡡ㘊మᙟᠺ࡞࠽࠷࡙ࡵࠉ53& ࡡ 3 ᮆ❻ࠉ( ᮆ❻࡛ )3&73& 㙈ࡡ ’ᮆ❻ࠉ’ᮆ❻ࡡ᪁ྡྷᛮ࡞
ᑊࡌࡾ㐽ᢝᛮ⏉࠷ࡆ࡛㔔さၡ㢗࡛ࡊ࡙ᣞᦤࡈࡿ࡙࠷ࡾࠊࡌࢂࡔࠉ53& ࡡ 3 ᮆ❻ഁ࡛ )3&73& ࡡ ’
ᮆ❻ഁ࡚┞Ⓩሲᇱᑊࢅᙟᠺࡌࡾ㏣ᖲ⾔㸝FQWLSFUFOOHO㸞ᆵ㘊మ࡛ࠉ3 ᮆ❻࡛ ’ᮆ❻ഁ࡚┞Ⓩ㔔㙈㘊
మࢅᙟᠺࡌࡾᖲ⾔㸝SFUFOOHO㸞ᆵ㘊మᏋᅹࡌࡾࠉ)3&73& 㘊మࡢᅸⓏ࡞㏣ᖲ⾔ᆵ㸝’ᮆ❻࣬’ᮆ❻㸞
Ꮽᏽ㘊మࢅᙟᠺࡌࡾࡡ࡞ᑊࡊࠉ53&࣬)3&73& 㘊మ࡚ࡢⱕᖱ㏣ᖲ⾔ᆵᏭᏽ࡚࠵ࡾࠉᖲ⾔ᆵ㘊మࡵ
Ẓ㍉ⓏᏭᏽ࡞Ꮛᅹࡊࠉࡐࡡㆉื⬗ࡢࡂࠉࡆࡿࣤࢲࢬࣤࢪฦᏄ࡛ࡊ࡙⏕࠷ࡾሔྙ࡞ኬࡀၡ㢗Ⅴ࡛
ࡊ࡙ᣞᦤࡈࡿ࡙࠷ࡾࠊࡱࡒࠉ࡞㏑ࡒ࣐ࢪ࣏ࢴࢲሲᇱࡡᏋᅹ࡞ࡻࡽ 53&࣬)3& 㘊మࡡᏭᏽᛮࡢኬࡀࡂ
ୖࡊ 9P ࡢ ࠤΥࡵୖࡌࡾࠉࡐࡿ࡚ࡵ )3&࣬)3& 㘊మࡡ 9P ࡻࡽ㧏ࡂࠉ࣐ࢪ࣏ࢴࢲࢅ᭯ࡌࡾ 53&࣬
)3& 㘊మࠉࣆ࣏ࣜࢴࢲ )3&࣬)3& 㘊మࡻࡽඁࡊ࡙⏍ᠺࡊ࡙ࡊࡱ࠹࡛࠷࠹ၡ㢗ࡵᢢ࠻࡙࠷ࡾࠊࡱࡒࠉ⌟
ᐁⓏࣤࢲࢬࣤࢪ 73& ฦᏄ࡛ࡊ࡙ࡡ㐲ᚺࢅ⩻࠻ࡾ࡛ࠉ53&࣬73& 㘊మ 73FVH- ࡡᇱ㈻࡛ࡽ㘊మ୯ࡡ
P73& ࡡࡲ 73FVH- ࡞ࡻࡽ㐽ᢝⓏ࡞ว᩷ࡈࡿࡾࡆ࡛࡞ࡻࡽࠉり፳㔖ࡡࣤࢲࢬࣤࢪฦᏄ࡞ࡻࡽຝᯕⓏᢒ
โຝᯕࢅⓆ⌟ࡌࡾࡆ࡛ịࡴࡼࡿࡾࠉ53&࣬73& 㘊మࡢ 73FVH- ࡡᇱ㈻࡛ࡢࡼ࠷ࡒࡴり፳Ⓩ࡞ࡢᶭ⬗
࡚ࡀࡍࠉP73& ࡛᭩Ꮥ㔖ㄵ㔖ࡡ 53& ࡡᢖᚪさࡆ࡛࡞ሒ࿈ࡈࡿ࡙࠷ࡾࠊ
ࡆࡡࡻ࠹ 53& ࡡ᭯ࡌࡾỀ⁈ᛮࠉᰶ㓗ヾㆉ᪁ྡྷᛮࡡၡ㢗よỬࠉ࠷⣵⬂⭯㏩㐛ᛮࡡྡྷ୕ࠉ㧏ࡌࡁࡾᏭ
ᏽᛮࡡ΅ࠉ73& ࡛ࡡ㘊మ 73FVH- ࡡᇱ㈻࡛ࡾࡆ࡛ࢅᐁ⌟ࡌࡾࡒࡴࠉࢲ࣭ࣔࢹ ࡞♟ࡊࡒ⛸ࠍࡡ
ಞ㣥 53& ሒ࿈ࡈࡿ࡙࠷ࡾ ࠊ⌟ᅹࡡಞ㣥 53& ◂✪ࡡ᪁ྡྷᛮࡢኬࡀࡂ௧ୖࡡ㸨ࡗ࡞ฦ㢦࡚ࡀࡾࠊ㸦㸞53&
ࡡỀ⁈ᛮࡡྡྷ୕ࡷ⣵⬂⭯㏩㐛ᛮࡷᰶᑻᅹࢅ┘Ⓩ࡛ࡊࡒ࢛ࣛࢥ࣋ࣈࢲࢺ㙈ࡷ )3&ࠉ⢶㙈࡛ࡡࣀࣇࣛ
ࢴࢺࠉ㸧㸞)3&73& 㙈ࡡ㒼ྡྷヾㆉᛮࡡྡྷ୕ࡡࡒࡴࠉSUHRUJFQL_H ࡈࡿࡒ㧏ḗᵋ㏸ࢅ᭯ࡌࡾ 53& ࡡྙᠺࠉ
㸨㸞㐿ఎᏄチ᩷࣬835V ᳠ฝ࣬ᰶ㓗≁␏Ⓩว᩷ࢅ┘Ⓩ࡛ࡊࡒ 53& ࡡᶭ⬗ࠉ࡚࠵ࡾࠊ
㸦㸞Ề⁈ᛮࡡྡྷ୕࡛ )3&73& ࡛ࡡࡻࡽᏭᏽ㘊మᙟᠺࢅ┘Ⓩ࡞ࠉ3࣐ࢿ࢙ࢲࣜࢡࣛࢨࣤࡡࢡࣛࢨ
ࣤṟᇱࢅࠉഁ㙈࡞ḿ㞹Ⲭࢅ᭯ࡌࡾࣛࢩࣤࡡሲᇱᛮ࣐ࢿ㓗࡞ንᥦࡊࡒㄇᑙమ 53& 㛜Ⓠิࡡṹ㝭
࡚ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉሲ⃨ᗐࡡẒ㍉Ⓩ㧏࠷⥾⾢ᾦ୯࡚ࡡ㘊మᙟᠺ⣌࡚࠵ࡾࡒࡴࠉ࡞ཬࡊ࡙㟴
㞹┞பష⏕࡞ࡻࡾኬᖕᏭᏽࡢሒ࿈ࡈࡿ࡙࠷࠷ࠊࡊ࠾ࡊ᭩㎾ࣜ࢟ࢼࣤࢅᑙථࡊࡒ 53& ㄇᑙమࡡྙ
ᠺሒ࿈ࡈࡿࠉ┞Ⓩ )3& ࡛ 53& ࡻࡽࡵࡻࡽᏭᏽ 㔔㙈㘊మࢅᙟᠺࡌࡾ࡛භ࡞ࠉ⣵⬂හࡡຝ
⋙Ⓩཱིࡽ㎲ࡲሒ࿈ࡈࡿࠉຝᯕⓏ⣵⬂හཱིࡽ㎲ࡲ᪁Ἢ࡛ࡊ࡙ᙽࡈࡿࡾ ࠊ
ୌ᪁ࠉᵕࠍᶭ⬗ᛮ࣋ࣈࢲࢺ࡛ 53& ࡛ࡡࣀࣇࣛࢴࢹࡢᰶ㓗࣋ࣈࢲࢺ々ྙᮞᩩ࡛ࡊ࡙ࡵ⮾࠵ࡾ
⣪ᮞ࡚࠵ࡽࠉ53& ࡡ 3 ᮆ❻ࡷ ( ᮆ❻࡚ᶭ⬗ᛮ࣋ࣈࢲࢺࢅ├⤎ྙࡊࡒࣀࣇࣛࢴࢺᆵฦᏄኣᩐሒ࿈ࡈࡿ
࡙࠷ࡾࠊ᭩㎾ᅖ┞Ἢ࡞ࡻࡾฦᒪᆵࡡ 53&࣋ࣈࢲࢺࣀࣇࣛࢴࢺࡡྙᠺἪሒ࿈ࡈࡿࡒ
㸝ࢲ࣭ࣔࢹ 㸞ࠊ
ᮇྙᠺἪ࡚ࡢࡒ࡛࠻ࡣ +PRHࣛࢩࣤࡡȜ࣐ࢿᇱࡡಕ㆜
ᇱ
࡛
ࡊ
࡙
ILPHWK^OILR]RFH^HORKH]^OLIHQHHWK^O
)IHᇱࢅ⏕࠷ࠉ+PRH ᇱ࡛ࡡ㐽ᢝⓏ⬲ಕ㆜࣬ᘇఘཬᚺࢅ⾔
O
Fmoc-HN
O
NH2OH
Rink
Fmoc-HN
CH2 4
NH-Dde
CH2
NH2
࠹ࡆ࡛࡞ࡻࡽᅖ┞Ἢ࡞ࡻࡽ⠾౼ࢸࣖࣜᆵ 53&࣋ࣈࢲ
ࢺ々ྙమྙᠺྊ⬗࡛ࡽࠉ᩺ࡊ࠷ᵋ㏸ࢅ᭯ࡌࡾ 53&࣋ࣈ
ࢲࢺࣀࣇࣛࢴࢹࡡྙᠺἪ࡛ࡊ࡙ᚃࡡᒈ㛜ᙽࡈࡿࡾࠊ
ୌ᪁ࠉኣᩐሒ࿈ࡡ࠵ࡾ├⤎ྙᆵࣀࣇࣛࢴࢹ࡞ࡗ࠷࡙
O
peptide
HN
CH2
NH
Chart 3 Synthetic scheme of !-Dde protected lysine.
6
ᖼࡗ࠾⣺ࡌࡾࠊ53& ࢅ⏕࠷ࡒ⣵⬂࡚ࣝ࣊ࣜᶭ⬗โᚒࡡࣈ࣭ࣞࢲ࡞㝷ࡊࠉኬࡀၡ㢗࡛ࡾࡡ࡞
ࡵ㏑ࡒ㟸ᖏ࡞࠷⣵⬂⭯㏩㐛ᛮ࡚࠵ࡾࠊ↋⣵⬂⣌࡚ࡢⰃይᢒโ࣬ὩᛮຝᯕࢅⓆ⌟ࡊ࡙࠷ࡾ⣌࡚ࡵ⣵
⬂⣌㐲⏕ࡊࡒሔྙ࡞ᶭ⬗Ⓠ⌟ふῼࡈࡿ࠷ཋᅄࡡୌࡗ࡛ࡊ࡙ࠉ⣵⬂හࡡཱི㎲㔖ᴗ❻࡞ᑛ࠷ࡆ
࡛ᣞᦤࡈࡿ࡙࠷ࡾࠊࡆࡡၡ㢗ࡡよỬἪࡡୌࡗ࡛ࡊ࡙ࠉ⣵⬂⭯㏩㐛ᛮ࣋ࣈࢲࢺ࡛ࡡ々ྙᥞ᱄ࡈࡿ࡙
࠷ࡾࠊ53& ࡢ࣋ࣈࢲࢺ㦭ࢅ᭯ࡌࡾࡒࡴࠉ3 ᮆ❻࠽ࡻࡦ ( ᮆ❻࡚ᶭ⬗ᛮ࢛ࣛࢥ࣋ࣈࢲࢺ࡛ࡡ々ྙࠉ
53&̺࣋ࣈࢲࢺ HKLPHUF ฦᏄࡡᒈ㛜ᐖ᪾࡚࠵ࡾࠊౚ࠻ࡣ⣵⬂⭯㏩㐛ᛮ࣋ࣈࢲࢺ࡚࠵ࡾ WUFQVSRUWFQ
ࡷ S&QWSࢅ├ࠉ࠵ࡾ࠷ࡢ࣭ࣛࣤ࢜ࢅࡊ࡙⤎ྙࡊࡒ 53& ㄇᑙమࡡሔྙࠉ↋ಞ㣥ࡡ 53& ࡚ࡢཱི㎲ධࡂ
㉫ࡆࡼ࠷⃨ᗐ࡚ࡵ㣍㌅Ⓩ⣵⬂හࡡཱི㎲ふῼࡈࡿࠉཱིࡽ㎲ࡱࡿࡒ 53& ࡞ࡻࡾࣤࢲࢬࣤࢪຝᯕ
࡞ࡻࡽࢰ࣭ࢣࢴࢹ P73& ࡡⓆ⌟ᢒโࢅᐁ⌟࡚ࡀࡾࡆ࡛ሒ࿈ࡈࡿࡒࠊࡱࡒࠉ53&̺ᶭ⬗ᛮ࣋ࣈࢲࢺ HKLPHUF
ฦᏄ࡛ࡊ࡙ࠉHP^H ࢝ࣤ㐿ఎᏄ࡞ᑊࡌࡾࣤࢲࢩ࣭ࣤ 53& ࡛ᰶ⛛⾔ᛮࢨࢡࢻࣜ࣋ࣈࢲࢺࡡ々ྙฦᏄྙ
ᠺࡈࡿࠉ53& ࡡᰶහ⃨ᗐࡡ୕࡛᪴ HP^HP73& ࡡ㌹ᢒโຝᯕሒ࿈ࡈࡿࡒ ࠊࡆࡿࡼࡡ◂✪ࡢᶭ⬗ᛮ࣋
ࣈࢲࢺ࡛ࡡ々ྙࠉ53& ࡡ⣵⬂㈻࠽ࡻࡦᰶ⛛⾔ࡡ㐽ᢝ㍲㏞࡞ຝᯕⓏ࡚࠵ࡾࡆ࡛ࢅ♟ࡊ࡙࠽ࡽࠉᶭ⬗
ᛮ࣋ࣈࢲࢺࡡ࣒࣭࣎ࣛࣚࢭ࡞ࡻࡾ⾉ᾦ୯࣬⣵⬂හฦよཬᚺࢅᢒโࡌࡾࡆ࡛ྊ⬗࡛ࡿࡣ᭯ຝ᪁Ἢㄵ
࡛ࡊ࡙ᙽࡈࡿࡾࠊࡱࡒࠉ53& ࡛ )3& ࡡࣀࣇࣛࢴࢹྙ∸ࡡྙᠺࡵሒ࿈ࡈࡿ࡙࠷ࡾࠊ࡞ࡵ㏑ࡒຝ⋙
ⓏࣤࢲࢬࣤࢪฦᏄࡡ㛜Ⓠࢅ┘Ⓩ࡞ 73FVH- Ὡᛮࢅ 53& ࡞ࡌࡾࡒࡴ )3& ࡡ ’ഁ࡛ ’ഁཫ᪁࡞ 53& ࢅ
⤎ྙࡊࡒ 53&̺)3&̺53& ࣒࢞ࣚฦᏄሒ࿈ࡈࡿ࡙࠷ࡾࠊ
ࡆࡡ࣒࢞ࣚฦᏄࡢ 53& ࡡ⤎ྙ࡞ࡻࡽࠉ
H]RQXHOHFVH
࡞ᑊࡌࡾ⪇ᛮࢅ⋋ᚋࡊࡗࡗࠉ73& ࡛ࡡ㘊మ 73FVH- ࡡᇱ㈻࡛ࡽ࠹ࡾࡆ࡛ሒ࿈ࡈࡿࠉࣤࢲࢬࣤࢪ 73&
ฦᏄ࡛ࡊ࡙᭯᭻ྙ∸࡚࠵ࡾ
ࠊࡈࡼ࡞ 53& ࡡ≁ᏽ⣵⬂ࡡࢰ࣭ࢣࢴࢹࢸࣛࣁ࣭ࣛࢅ┘Ⓩ࡛ࡊ࡙⢶㙈
࡛ࣀࣇࣛࢴࢹࡊࡒㄇᑙమࡡྙᠺࡵሒ࿈ࡈࡿ࡙࠷ࡾࠊ53& 㦭࡞ࢬࣛࣤࡷࢪ࢛ࣝࢼࣤࡐࡊ࡙ࣛࢩࣤࡷ
ࢪࣂࣚ࢟ࣤࢅᑙථࡊࠉࡐࡿࡑࡿ 4ࢡࣛࢤࢨࣜࠉ(ࢡࣛࢤࢨࣜࠉ3ࢡࣛࢤࢨࣜࡊ⢶㙈ࢅᑙථࡊࡒ 73&
࡛ࡡ㘊మᏭᏽᛮࡢୖࡌࡾࡵࡡࡡࠉ3ࢬࢲࣜࢡࣜࢤࢦ࣐ࣤࢅᑙථࡊࡒ 53& ࡢ⫚⮒≁␏Ⓩ࡞ཱིࡽ㎲ࡱ
ࡿࡾࡆ࡛ሒ࿈ࡈࡿ࡙࠷ࡾࠊ⫚⮒ࡡ≁␏Ⓩࢸࣛࣁ࣭ࣛࡢẒ㍉Ⓩᐖ᪾࡚࠵ࡽ┘᩺ࡊࡈࡢ࠷ࠉ⢶㙈
࡛ࡡࣀࣇࣛࢴࢹ࡛࠷࠹Ⅴ࡚⮾ࡂࠉᚃࡡ㐅ᒈ࡞⮾ᣚࡒࡿࡾ ࠊ
㸧㸞)3&73& 㙈ࡡ㒼ྡྷヾㆉᛮࡡྡྷ୕ࡡࡒࡴࠉ53& ㄇᑙమ࡞ SUHRUJFQL_H ࡈࡿࡒ㧏ḗᵋ㏸ࡡ᭯ຝ࡛
ࡡ௫ㄕ࡞ᇱࡘࡀࠉᵕࠍ 53& ㄇᑙమྙᠺ✒ᴗⓏ࡞᳠ゞࡈࡿ࡙࠷ࡾࠊරమⓏ࡞ࡢࢨࢠࣞ࣋ࣤࢰࣤ⎌
ࢨࢠࣞࣉ࢞ࢦࣤ⎌
ࠉࣈࣞࣛࣜࣅ࣋ࢤࣛࣜ㦭
ࠉࣅࣞࣛࢩࢿ㦭
ࡷ
ࢅ᭯ࡌࡾ 53& ㄇᑙమሒ࿈
ࡈࡿ࡙࠷ࡾࠊ☔࠾࡞ࠉ㏣ᖲ⾔ᆵ㘊మࡡᏭᏽᛮᖲ⾔ᆵ㘊మࡻࡽ㧏ࡂࡾ⣌ࡢሒ࿈ࡈࡿ࡙࠷ࡾࠉ㘊మᏭ
ᏽᛮࡢ࠷ࡍࡿࡡ⣌࡚ࡵ 53&࣬)3& 㘊మࡡᏭᏽᛮࡻࡽࡵ΅ࡌࡾലྡྷ࡞࠵ࡽࠉࡱࡓࡱࡓ᳠ゞࡡఴᆀ࠵ࡾࠊ
ࣅࣞࣛࢩࢿ㦭ࢅ᭯ࡌࡾ 53& ㄇᑙమ࡚ࡢ㏳ᖏࡡ 53& ࡛ࡢ㏣࡞ )3& ࡛ࡡ㘊మ 73& 㘊మࡻࡽᏭᏽ࡚࠵ࡾࡆ
࡛ሒ࿈ࡈࡿ࡙࠷ࡾࠉSRO^&࡛ࡡ㘊ᙟᠺ⣌࡚ࡡ⤎ᯕ࡞ᇱࡘࡀレ⣵᳠ゞᚪさ࡚࠵ࢀ࠹ࠊ㙈㦭
࡞ⰶ㤮⎌ࢅᑙථࡊࡒ 53& ㄇᑙమࡡྙᠺࡵ᳠ゞࡈࡿ࡙࠷ࡾࠊࡆࡡ࣓ࢸ࡚ࣜࡵฦᏄහ࡚ࡡࢪࢰࢴ࢞ࣤࢡ┞ப
ష⏕࣬Ề┞பష⏕࡞ࡻࡾ㧏ḗᵋ㏸ࡡࢅ┘Ⓩ࡞ฦᏄスゝࡈࡿࠉFQWLSFUFOOHO ᆵࡡ 73& ࡛ࡡ㘊మᏭ
ᏽ࡞ᙟᠺࡈࡿࡾࡆ࡛ࢅሒ࿈ࡊ࡙࠷ࡾ༎ฦỀ⁈ᛮࢅ♟ࡌࡡ࠾ၡ࡚࠵ࡾ ࠊࡱࡒࠉ᭩㎾Șࣉࣛࢴࢠࢪ
ᵋ㏸ࢅ᭯ࡌࡾ 53& ㄇᑙమࡡྙᠺሒ࿈ࡈࡿࠉSFUFOOHO ᆵ㘊మ FQWLSFUFOOHO ᆵ㘊మࡻࡽᏭᏽ࡚࠵ࡾࡆ࡛
ሒ࿈ࡈࡿ࡙࠷ࡾࠉ࡛ྜྷᵕ࡞ࡐࡡᏭᏽᛮࡡᕣ␏ࡢ༎ฦ࡛ࡢゕ࠻ࡍࠉࡱࡒ㘊మᏭᏽᛮࡵ 53& 㘊మࡡࡐ
ࡿ࡛Ẓ㍉ࡊ࡙࠾ࡽୖࡊ࡙࠷ࡾࠊሲᇱ㒂ࡡࢪ࣭࣋ࢦ࣭࡛ࡊ࡙࢙࣭ࢷࣜ⤎ྙࢅᑙථࡊ㌶ᛮࢅࡊ࡙
࠷ࡾࡵࡡࡡ㙈㦭Ș࣋ࣈࢲࢺ㦭࡚࠵ࡾࡒࡴ࠾ࡽ๙├Șࣉࣛࢴࢠࢪᵋ㏸ࢅᣚࡔࠉࣉࣛࢴࢠࢪ
7
ࣅࢴࢲ࡛ࢽࢠ࢛ࣝࢲࢺࡡሲᇱ㛣㝰␏ࡾࡆ࡛ࡵ࠵ࡽࠉධሲᇱᑊᙟᠺࡈࡿ࡙࠷࠷ྊ⬗ᛮࡵ࠵ࡽࠉ
ᚃᵋ㏸᭩㐲ᚪさ࡛ࡵ⩻࠻ࡼࡿࡾ ࠊ
ࡱࡒỀ⁈ᛮࡡྡྷ୕࡛භ࡞࣐ࢪ࣏ࢴࢲ)3&࡛ࡡ㘊మᏭᏽᛮࡡ㐲ḿࢅ┘Ⓩ࡞ࠉᏱᡖࡼࡢ㌶࢙࣭ࢷࣜ⤎
ྙࢅ㙈㦭࡞ᑙථࡊࡒ࢛࢞ࢨ࣋ࣈࢲࢺᰶ㓗㸝453&㸞ࢅスゝ࣬ྙᠺࡊ࡙࠷ࡾ ࠊ453&ࡢⰃይỀ⁈ᛮࢅ
♟ࡌ࡛භ࡞ࠉࣈࣛࣤሲᇱࢅ᭯ࡌࡾ453&ࡢ53&࣬)3&㘊మࡡᏭᏽᛮ࡞┞ᙔࡌࡾᏭᏽ㘊మࢅᙟᠺࡌࡾ࡛භ࡞ࠉ
㙈ࡡ㌶ᛮ࡞㉫ᅄࡌࡾ࢙ࣤࢹࣞࣅ࣭ࣆࢠࢰ࣭ࡡᐞ࡞ࡻࡽ㟸ᖏ࡞ࢨ࣭ࣔࣈ⼝よ᭜⥲ࢅ♟ࡊࠉ≻࠷
Ὼᗐ⠂ᅑ࡚㘊మࡡᙟᠺ࣬よ㞫ྊ⬗࡛ࡾඁࡿࡒ≁ᚡࢅ♟ࡌࠊ
㸨㸞53& ࡡᶭ⬗࡞㛭ࡊ࡙ࡢ㟸ᖏ࡞ኣࡂࡡ◂✪ሒ࿈ࡈࡿ࡙࠽ࡽࠉ⣤㟻ࡡ㛭౿ࡵ࠵ࡽධ㒂ࢅ⣺ࡌࡾࡆ࡛
ࡢᗇྊ⬗࡚࠵ࡾࡡ࡚ࠉࡆࡆ࡚ࡢᖼࡗ࠾᭩㎾ࡡࢹࣅࢴࢠࢪࢅ⣺ࡊࡒ࠷ࠊ
ᶭ⬗ᛮ 53& ࡡ㐲⏕᳠ゞࡈࡿ࡙࠷ࡾ⌟ᅹ᭩ࡵ⢥ງⓏ࡞◂✪ࡈࡿ࡙࠷ࡾฦ㔕ࡡୌࡗ࡛ࡊ࡙ࠉ835V ᳠ฝࢅ
୯ᚨ࡛ࡊࡒ㐿ఎᏄチ᩷࠵ࡾࠊ㐿ఎᏄチ᩷ࡢ⑄࣬⑄ᝀཋᅄ㐿ఎᏄࡡ㐿ఎᏄ㒼าࡡ┞㐢ࠉ835V ࢅࢰ
࣭ࢣࢴࢹ࡛ࡊ࡙ࡐࡡን␏㐿ఎᏄ࡞ሲᇱ㒼า≁␏Ⓩ࡞⤎ྙࡌࡾࡆ࡛ᇱᮇ࡚࠵ࡽࠉ࠷࠾࡞ሲᇱ㒼า≁␏Ⓩ
࡞ࢰ࣭ࢣࢴࢹ )3&73& ࡞⤎ྙࡌࡾ࠾㔔さၡ㢗࡛ࡾࠊῼᏽ᪁Ἢࡢ )3& ࢲࢴࣈ࡛ࡻࡣࡿࡾᇱᯀ୕࡞㐿
ఎᏄチ᩷⏕᳠ฝモⷾࢅᅖᏽࡊኣモᩩྜྷ᳠ฝࡌࡾ᪁Ἢ࡛ࠉᆍୌ⁈ᾦ⣌࡚᳠ฝࢅ⾔࠹ᆍୌ⁈ᾦἪ࡞ኬื
ࡈࡿࡾࠊ࠷ࡍࡿࡡ⣌࡚ࡵ᳠ฝ᪁Ἢࡡᇱᮇࡢ ࠤ2 ࡡ㧏࠷᳠ฝវᗐࢅ᭯ࡌࡾ⺧ක᳠ฝἪ⏕ࡈࡿ࡙࠽ࡽࠉ
≁࡞ ⛸㢦ࡡ⺧කⰅ⣪㛣ࠉ⺧කⰅ⣪࡛ᾐක㛣࡚ࡡ࢙ࢾ࣭ࣜ࢟⛛ິࢅฺ⏕ࡊࡒ⺧කභ㫾࢙ࢾ࣭ࣜ࢟⛛ິ
㸝+7*9㸞ࠉってⓏ࡞ࡵ᳠ฝྊ⬗࡚࠵ࡾࡆ࡛࠾ࡼ᭩㎾Ἰ┘ࡈࡿ࡙࠷ࡾࠊᇱᮇⓏ᳠ฝཋ⌦ࢅࢲ࣭ࣔࢹ ࡞♟ࡊࡒࠊ࣓࣭ࣝ࢞ࣖࣚࣄ࣭ࢤ࡛ࣤྞࡄࡼࡿࡒ 3 ᮆ❻࡛ ( ᮆ❻ཫ᪁࡞ທ㉫ἴ㛏ࡼࡦ࡞Ⓠකἴ㛏␏
ࡾ ⛸㢦ࡡ⺧කⰅ⣪࠵ࡾ࠷ࡢ⺧කⰅ⣪࡛ᾐකࢅ⤎ྙࡊࡒ⮤ᕤ┞Ⓩ 53& ࢅྙᠺࡊࠉࢰ࣭ࢣࢴࢹ )3&
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Chart 4 Schematic drawing of molecular beacon PNA.
࡛Ίྙࡌࡾ
ࠊ⺧කᵾㆉ )3& ࣈ࣭ࣞࣇ࡚࠵ࡾ⮤ᕤ┞Ⓩ 53& ࡢࢰ࣭ࢣࢴࢹ࡛⤎ྙࡊ࡙࠷࠷≟ឺ࡚ࡢ ⛸ࡡ⺧කⰅ⣪ࠉ࠵ࡾ࠷ࡢ⺧කⰅ⣪࡛ᾐකࡢ㎾ࡌࡾࡻ࠹࡞スゝࡈࡿ࡙࠷ࡾࡡ࡚ࠉ ⛸ࡡ⺧කⰅ⣪ࢅ᭯ࡌ
8
ࡾ⣌࡚ࡢ㛏ἴ㛏⺧කⰅ⣪࠾ࡼࡡⓆකࠉᾐක࡛ࡡ⤄ྙࡎࡡ⣌࡚ࡢᾐකࡈࡿ⺧කࢅⓆࡊ࠷ࠊ┞Ⓩࢦ
ࣤࣈࣜ )3& ࢅ῟ຊࡌࡾ࡛ࠉ53&࣬)3& 㔔㙈ࢅᙟᠺࡌࡾࡆ࡛࡞ࡻࡽ⺧කⰅ⣪㛣㊝㞫ࠉⰅ⣪࣬ᾐක㊝㞫
㞫ࡿࠉ ⛸㢦ࡡ⺧කⰅ⣪࠾ࡼࡡⓆකࡼࡦ࡞ᾐක≟ឺよࡄⓆකふῼࡈࡿࢰ࣭ࢣࢴࢹ )3& ࡡᏋᅹ⺧ක
Ⰵㄢንࡼࡦ࡞Ⓠක࡞ࡻࡽ᳠ฝྊ⬗࡛ࡾࠊࡆࡡ࣓࣭ࣝ࢞ࣖࣚࣄ࣭ࢤࣤࡢᙔิ⮤ᕤ┞Ⓩ )3& ࢅ⏕࠷
ࡒ⣌ሒ࿈ࡈࡿࡒࠉ㓕⣪ฦよ࡞ࡻࡾ්⌟ᛮࡡ㞬ࡊࡈᣞᦤࡈࡿࠉᏕⓏ࣬㓕⣪Ⓩ࡞Ꮽᏽ 53&
ࢅ⏕࠷ࡾࡆ࡛࡞ࡻࡽ්⌟ᛮ☔ಕࡈࡿࡾ࡛භ࡞ࠉ53& ࡡ㧏࠷⤎ྙᏽᩐ࡞ᇱࡘࡀ⃨ᗐ࡚ࡵ )3& ࢅ㧏វᗐ᳠
ฝ࡚ࡀࠉ)3& 㧏ḗᵋ㏸ࢅཱིࡽࡘࡼ࠷ሲ⃨ᗐ࡚ࡵᏭᏽ࡞㔔㙈㘊మࢅᙟᠺࡌࡾࡆ࡛࠾ࡼಘ㢏ࡡฝᮮࡾ᳠
ฝἪ࡛ࡊ࡙☔❟ࡈࡿࡗࡗ࠵ࡾࠊࡱࡒࠉ53& ࡡ㧏࠷ሲᇱ㒼า㐽ᢝᛮ࡛㓕⣪⪇ᛮࢅฺ⏕ࡊ 53& ࡛ 3XHOHFVH8
ࢅ⤄ྙࡎࠉ53&࣬)3& 㘊మ࡞≁␏Ⓩ࡞⤎ྙࡌࡾ )L8( ⺧කⰅ⣪ࢅฺ⏕ࡊࡒ㧏វᗐ᳠ฝࢨࢪࢷ࣑ᑚᐋᒜࡼ
࡞ࡻࡽሒ࿈ࡈࡿࠉἸ┘ࡈࡿ࡙࠷ࡾ ࠊࡱࡒ ௧୕ࡡ࢝ࣤ⣵⬂࡚ን␏ሒ࿈ࡈࡿ࡙࠷ࡾࢨࢹࢨࣤ㸝(㸞Ѝ
࣒ࢲࣜࢨࢹࢨࣤ㸝P(㸞ࡡን␏ࢅ 53&IV)3& 㘊మࡡᙟᠺࡌࡾ 5ORRS ࡛โ㝀㓕⣪ࢅ⤄ࡲྙࢂࡎࠉ+7*9 ࢅ
⏕࠷࡙᳠ฝࡌࡾ᪁Ἢሒ࿈ࡈࡿࠉᚉᮮᅏ㞬࡚࠵ࡖࡒ P( ࡡ᳠ฝ᪁Ἢ࡛ࡊ࡙Ἰ┘ࡈࡿ࡙࠷ࡾ ࠊ
53& ࡡ࣓ࢿ࣏Ѹࡢୌᕰ㈅ࡵࡈࡿᅖ┞ྙᠺࡷ⮤ິྙᠺᶭࢅ⏕࠷࡙Ẓ㍉Ⓩ⠾౼࡞࢛ࣛࢥ࣏࣭ྙᠺྊ⬗࡚ࠉ
ࡈࡼ࡞౪㢏ྙᠺ࡚ࡵ┘Ⓩ࢛ࣛࢥ࣏࣭ࢅථᡥࡌࡾࡆ࡛ࡵྊ⬗࡞ࡵࡖ࡙࠷ࡒࠊࡆࡡࡒࡴࠉᏕ⩽ࡓࡄ࡚ࡢ
ࡂࠉ⏍Ꮥࠉ⏍∸ᕝᏕࡐࡊ࡙ฦᏄ⏍∸Ꮥࢅᑍ㛓࡛ࡌࡾ◂✪⩽ࡵ࢜ࢪࢰ࣑ྙᠺࡡ )3&73& ࡛ྜྷᵕ࡞ 53&
ࢅ⏕࠷ࡒ◂✪࡞ཱིࡽ⤄ࡴࡾ⎌ሾ࡛ࡽࠉ㸦㸞㐿ఎᏄ )3& ࠾ࡼ SUHP73& ࡡ㌹㐛⛤ࡡᢒโࠉ㸧㸞P73& ࠾
ࡼࡡࢰࣤࣂࢠ㈻ྙᠺࡡ⩳ゼ㐛⛤ࡡᢒโࠉ㸨㸞㌹㡷ᇡࣤࢲࢬࣤࢪ㙈 )3& ࡞⤎ྙࡌࡾࡆ࡛࡞ࡻࡾ㌹
Ὡᛮࡡቌ㐅ࠉࡡ 53& ࢅ⏕࠷ࡒ㐿ఎሒⓆ⌟โᚒ࡞㛭ࡌࡾኣᩐࡡሒ࿈ࡵࡈࡿ࡙࠷ࡾࠊ
᭩ᚃ࡞ )3& ࢅࢷࣤࣈ࣭ࣝࢹ࡛ࡌࡾ 53& ㄇᑙమࡡࢷࣤࣈ࣭ࣝࢹ࢛ࣛࢥ࣏࣭ሒ࿈ࡈࡿࡒࠊ53& 㔖మࡡ
( ᮆ❻ࢅࣜࢸࣃࢺ࡛ࡊࠉ53& 㔖మ࡛┞Ⓩ㒼าࢅ ᅂࡡ⧖ࡽ㏁ࡊ㒼าࢅ᭯ࡌࡾ )3& Ꮛᅹୖࢨࢴࣆሲ
ᇱࡡᙟᠺࠉ3F'-(3 ࡞ࡻࡾ㑇ඔ࡞ࡻࡽຝ⋙Ⓩ 53& ྙᠺࢅሒ࿈ࡊ࡙࠷ࡾࠊ㔔㙈ᙟᠺ࡞ฺ᭯ 53& 㔖మ
ࢅ⏕࠷࡙࠽ࡽࠉຝᯕⓏሲᇱᑊᙟᠺ࡛ࡐࡿ࡞ᘤࡀ⤾ࡂᘇఘཬᚺࡢ༎ฦ࡞ྊ⬗࡚࠵ࡽࠉ┘᩺ࡊࡈࡢ
࠷ࠉ)3& ࠾ࡼ 53& ࡡሒ㌹࡛࠷࠹ふⅤ࠾ࡼࡢ⮾ࡂࠉᚃ 53& ࣓ࢿ࣏Ѹࢅ⏕࠷ࡒ⣌࡚ࢷࣤࣈ࣭ࣝ
ࢹ࢛ࣛࢥ࣏࣭ࡡᐁ⌟ᙽࡈࡿࡾ ࠊ
௧୕ᖼࡗ࠾ࡡౚࢅᣪࡅࡼ 53& ◂✪ࡡ⌟≟ࢅ⣺ࡊࡒࠉరࡼ࠾ࡡᙟ࡚ 53& ࡷࡐࡡㄇᑙమࢅ⏕࠷ࡒ
◂✪ࡢᣞᩐ㛭ᩐⓏ࡞ቌኬࡊ࡙࠽ࡽࠉ⌟ᅹ࡚ࡢ㸝3LHOVHQ ࡼ᭩ิࡡㄵᩝࡡ୯࡚ࡵ⏕࠷࡙࠷ࡾࡡࡓ㸞53&
ࡢ࣐࣎ࣛࢺᰶ㓗㸝5RO^FPLIH3XHOHLH&HLI㸞࡛ࡊ࡙ࠉኣࡂࡡெᕝᰶ㓗ࡡ␆⛘࡛ࡊ࡙⏕࠷ࡼࡿࡾࡆ࡛ࡵ
࠵ࡾࠊ
ๆ⃥ᚺ➽ᛮெᕝᰶ㓗ࡡᵋ⠇
ࡆࡡࡻ࠹࡞ 53& ࡢᩐࠍࡡ≁ᛮࢅ᭯ࡊࠉḖⅤࢅග᭱ࡌࡾ 53& ㄇᑙమࡡ◂✪ࡵ⢥ງⓏ࡞⾔ࢂࡿ࡙࠷ࡾࠊࡊ
࠾ࡊࠉᚉᮮሒ࿈ࡈࡿ࡙࠷ࡾ 53& ࡵྱࡴࡒ࡛ࢆࡌ࡙ࡡᰶ㓗࣓ࢸࣜ࣬ெᕝᰶ㓗ࡢࠉࢰ࣭ࢣࢴࢹ )3&࣬73&
࡛࠷࠾࡞ᙁࡂ⤎ྙࡌࡾ࠾ࠉ9P ࡡ㧏࠷㘊మࢅᙟᠺࡌࡾ࠾ࢅ┘ᵾ࡞スゝ࣬ྙᠺ࡞ງἸࡿ࡙ࡀࡒࠊ
ౚ࠻ࡣࣤࢲࢬࣤࢪฦᏄ࡞㛭ࡊ࡙ࡢࠉP73& ࡛ࡡᏭᏽ㘊మᙟᠺ࡞ࡻࡾ㐿ఎሒࡡⓆ⌟ᢒโࡡࡲࢅ┘Ⓩ
࡛ࡊ࡙࠷ࡾࠊࡈࡼ࡞⛸ࠍࡡ༈⏕ᮞᩩࠉ≁࡞ 835V 㐿ఎᏄチ᩷ᮞᩩࡡᒈ㛜ࢅౚ࡛ࡊࡒሔྙࡵࠉࢰ࣭ࢣ
ࢴࢹ )3&࣬73& ࡛⤎ྙࡊࠉ࠷࠾࡞㧏វᗐ࡞᳠ฝࡌࡾ࠾ၡ㢗࡚ࠉୌ᪁ྡྷࡡ㘊మᙟᠺࢅ┘ᵾ࡞◂✪ᒈ㛜ࡈ
ࡿ࡙࠷ࡾࠊࡓࠉ༟⣟࣬ୌ᪁ྡྷ㘊మᙟᠺࡓࡄ࡚ࡢࡂࠉࢰ࣭ࢣࢴࢹ )3&࣬73& ࡛ࡡ㘊మᙟᠺ࣬よ㞫ࡡྊ㏣
Ⓩโᚒࠉࡌࢂࡔヾㆉ࣬㘊ᙟᠺ⬗ࡡአ㒂ᅄᏄ࣬አ㒂ๆ⃥࡞ࡻࡾ⮤ᅹ࡚ྊ㏣Ⓩโᚒྊ⬗ெᕝᰶ㓗㛜
9
Ⓠࡈࡿࡿࡣࠉࣤࢲࢬࣤࢪ⒢࡞࠽࠷࡙ࡵᢒโຝᯕࡡࡲࡼࡍࠉ㐿ఎሒࡡⓆ⌟࣬ᢒโࡡ⮤ᅹ࡚ྊ㏣Ⓩ
ࢤࣤࢹ࣭ࣞࣜྊ⬗࡞ࡾࡆ࡛ᙽ࡚ࡀࠉࣤࢲࢬࣤࢪฦᏄࡡ㐲⏕⠂ᅑࡡ㣍㌅Ⓩᣉኬྊ⬗࡛ࡾࠊ
ࡱࡒࠉᰶ㓗ࡡฦᏄヾㆉ࡞ᇱࡘࡂᵕࠍᶭ⬗ᮞᩩࡡ㐲⏕࡞࠽࠷࡙ࡵࠉᮞᩩᶭ⬗ࡡአ㒂ๆ⃥࣬አ㒂ᅄᏄ࡞
ࡻࡾ RQRII ࢪࢴࢲࣤࢡโᚒᐁ⌟࡚ࡀࠉெᕝᰶ㓗ࡡᮞᩩᒈ㛜ࡵኬᖕ࡞ຊ㏷ࡈࡿࡾࡆ࡛ᙽࡈࡿࡾ㸝ࢲ
࣭ࣔࢹ 㸞ࠊࡆࡿࢅᐁ⌟ࡌࡾࡒࡴ࡞ࡢࠉአ㒂࠾ࡼࡡๆ⃥ࡷአ㒂ᅄᏄ࡞ࡻࡾᰶ㓗ヾㆉࡡྊ㏣Ⓩโᚒࠉࢰ࣭ࢣ
ࢴࢹ࡛ࡡ㘊మᙟᠺ࣬よ㞫โᚒᚪさ࡛ࡾࠊ᭩㎾ࠉᑚᐋᒜ࣬ὰ࡞ࡻࡖ࡙ࣛࣤ㓗ࢩ࢙ࢪࢷࣜ㒂న࡞ࢯ
࣊ࣤࢭࣤㄇᑙమࢅᑙථࡊࡒಞ㣥 )3& ࢅ⏕࠷࡙ࠉ⣰አක࣬ྊっක↯ᑏ࡞ࡻࡾሲᇱ㒼า≁␏Ⓩ㔔㙈࠽ࡻ
ࡦ㔔㙈㘊మᙟᠺ࣬よ㞫โᚒࡼࡦ࡞ࠉක࡞ࡻࡾ࣒࣭࣎ࣛࣚࢭὩᛮโᚒྊ⬗࡛ࡾ࡛࠷࠹⮾࠷ㄵ
ᩝሒ࿈
ࡈࡿࡒࠉࡆࡿ௧አ࡞ࡢአ㒂ᅄᏄ࣬አ㒂ๆ⃥࡞ࡻࡾྊ㏣Ⓩᰶ㓗ヾㆉᶭ⬗ࢅ᭯ࡌࡾᰶ㓗࣓ࢸ
ࣜ࡞㛭ࡌࡾሒ࿈ࠉࡼࡦ࡞ࡐࡡ᪁Ἢㄵ࡞㛭ࡌࡾሒ࿈ࡈ࠻࡛ࢆぜ࠵ࡒࡼ࠷ࠊ
ࡆࡡࡻ࠹⫴ᬊࢅ㊻ࡱ࠻ࠉᠻࠍࡢḗୠࡡࣤࢲࢬࣤࢪฦᏄ࡛ࡊ࡙ࢰ࣭ࢣࢴࢹᰶ㓗࡛Ꮽᏽ㘊మࢅᙟ
ᠺࡌࡾ࡛࡛ࡵ࡞ࠉ⣵⬂හ⎌ሾࡡአ㒂ᅄᏄ࣬አ㒂ๆ⃥࡞ࡻࡾྊ㏣Ⓩ㘊మᙟᠺ࣬よ㞫โᚒྊ⬗ெᕝ
ᰶ㓗㛜Ⓠ࡞ཱིࡽ⤄ࢆ࡚࠷ࡾࠊ࡞ࡵ㏑ࡒྊ㏣Ⓩሲᇱ㒂ヾㆉโᚒ࡞㛭ࡊ࡙ࡢࠉࡆࡿࡱ࡚ࡐࡡ᪁Ἢㄵ
ࡈ࠻ᥞ᱄ࡈࡿ࡙࠷࠾ࡖࡒࠊ
ᠻࠍࡢࠉᰶ㓗ࡡヾㆉ㐛⛤࡞࠽࠷࡙ࡢሲᇱ㒂ࡡ㒼ྡྷ㔔さ࡚࠵ࡽࠉຝᯕⓏሲᇱヾㆉ࡞ࡢࣅ࣐ࣛࢩࣤሲ
ᇱ࡚ࡢ న࢜ࣜ࣍ࢼࣜᇱࠉࣈࣛࣤሲᇱࡡሔྙࡢࣅ࣐ࣛࢩࣤ㒂⢶㒂ࡡཬᑊഁ࡞Ꮛᅹࡌࡾ FQWL 㒼ྡྷࢅඁ
ࡌࡾᚪさ࠵ࡽࠉ㏣ࡡ V^Q 㒼ྡྷࡢሲᇱヾㆉ࡞࡛ࡖฺ࡙࡚᭯ࡾࡆ࡛ࠉFQWLѸ V^Q ሲᇱ㒂㒼ྡྷࡢ⢶㒂ࡡࢤ
࣒࣭ࣤ࣌ࢨࣘࣤ࡞ࡻࡽᙫ㡢ࢅུࡄࡾࡆ࡛࡞Ἰ┘ࡊࡒࠊࡌࢂࡔࠉ⢶㒂 ’’Ề㓗ᇱࢅ᭯ࡌࡾᰶ㓗ヾㆉฦ
አ㒂ᅄᏄ
Ề⣪⤎ྙRQЍRII
PRNA
࣌㓗㢦
RNA
S-࣬⢶⃨ᗐ࣬Ὼᗐ࣬HWH
573&73&㔔㙈
O
HN
H
N
O
N
O
O
H NH
UN H
HO OH
N
N
O
anti
O
HO
N
NA
N
O
O
O
ᰶ㓗ヾㆉ࡞ᇱࡘࡂᶭ⬗Ⓠ⌟4Q
࣬ࣤࢲࢬࣤࢪ73&࣬
ࠈࣤࢲࢩѸ ࣤᶭ⬗Ⓠ⌟
࣬㐿ఎᏄチ᩷RQ
࣬)3&ࢲࢴࣈRQ
࣬ᰶ㓗࣬ࢰࣤࣂࢠ々ྙమᙟᠺ
࣬㉰ฦᏄᙟᠺ
H
O
O
N
syn
O
O O
B
HO OH
NH2 N
NA N
N
U
O
O
P
HN
HN
O
HO
O
O
O
O
P
O
ᰶ㓗ヾㆉ࡞ᇱࡘࡂᶭ⬗Ⓠ⌟4II
࣬ࣤࢲࢬࣤࢪ73&࣬
ࠈࣤࢲࢩѸ ࣤᶭ⬗RII
࣬㐿ఎᏄチ᩷RII
࣬)3&ࢲࢴࣈRII
࣬ᰶ㓗࣬ࢰࣤࣂࢠ々ྙమよ㞫
࣬㉰ฦᏄよ㞫
Chart 5 The schematic drawing of reversible control of recognition and complexation behavior of PRNA by borax.
Ꮔ࡞࠽࠷࡙ࠉአ㒂ᅄᏄ࡞ࡻࡽ⢶㒂ࢤ࣒࣭ࣤ࣌ࢨࣘࣤንࢅㄇ㉫࡚ࡀࡿࡣࠉሲᇱ㒂ࡡ㒼ྡྷโᚒྊ⬗࡛
ࡽࠉࡐࡡ⤎ᯕ࡛ࡊ࡙ヾㆉโᚒᙽ࡚ࡀࡾ㸝ࢲ࣭ࣔࢹ 㸞ࠊࡆࡡふⅤ࠾ࡼࠉአ㒂ᅄᏄ࡞ࡻࡾሲᇱ㒂㒼ྡྷโ
10
ᚒࠉᰶ㓗ヾㆉโᚒ࡞ཱིࡽ⤄ࢆࡓࠊࡐࡊ࡙ 73& ࢅᰶ㓗ヾ
(a) Phosphate buffer
ㆉ 㒂 న ࡛ ࡊ ࡙ ᑙ ථ ࡊ ࡒ ࣋ ࣈ ࢲ ࢺ ࣛ ࣍ ᰶ 㓗 㸝 SHSWLIH
H5 H1'
H6
1
ULGRQXHOHLHFHLI 573&㸞࡛ྞࡄࡒ᩺ࡊ࠷࢜ࢷࢥࣛ
H3'H4'
H2'
H NMR
H5'
࣭ࡡெᕝᰶ㓗ࢅスゝ࣬ྙᠺࡊࡒ ࠊ73& ࡢࠉ)3& ࡞Ẓ
㍉ࡊ࡙ P73&ࠉW73&ࠉU73& ࡞⾪ࡈࡿࡾࡻ࠹࡞ᵕࠍ
㧏ḗᵋ㏸࡛㧏ᗐᶭ⬗ࢅⓆ⌟ࡌࡾࡆ࡛▩ࡼࡿ࡙࠷
7
O
ࡾࠊࡈࡼ࡞ 2J㒼నࡊࡒ 73& 㔘ᒌ㓕⣪࡛ࡊ࡙
H2N H H
H 5' H
O
ష⏕ࡌࡾࣛ࣍ࢧ࣑ࡷว᩷Ὡᛮࢅ᭯ࡌࡾ 73&L ࡵⓆぜࡈ
4'
H HO
NH
6
N
O
4
3 ppm
H2'
NOE
H3'
H5'
HH
OH
ࡿࠉࣈࢰ࣏࣭ࡡᚺ⏕ࢅࡢࡋࡴ 73& ࡡ♟ࡌ㧏ᗐᶭ
7
⬗࡞Ἰ┘㞗ࡱࡖ࡙࠷ࡾࠊࡊ࠾ࡊࠉ73& ࡢ ’Ề㓗ᇱ࡞
5
1'
2'
3'
6
H5 H1'
3
H 5
(b) Borate buffer
6
H5
5
4
3 ppm
H6
ᇱࡘࡂ㦭ᵋ㏸Ꮽᏽᛮࢅ♟ࡊࠉᐖ᪾࡞ຊỀฦよࢅུ
1
H1'
H NMR
H5'
H3'H4'
H2'
ࡄὩࡊ࡙ࡊࡱ࠹ࡡᮇ㈻Ⓩ࡚็ၡ㢗ࡵᢢ࠻࡙
࠷ࡾࠊࡆࡡࡒࡴࠉ73& ࢅᵋᠺ༟న࡛ࡌࡾெᕝᰶ㓗ࡢ⮾
ᣚࡒࡿ࡙࠷ࡾࠉࡆࡿࡱ࡚ 73& ࢅ᭯ࡌࡾᰶ㓗࣓ࢸࣜ
O
3
HN
H
O
H
H NH
N
5' H O H
ྙ∸ࡢ࡛ࢆሒ࿈ࡈࡿ࡙࠷࠷ࠊࡆࡡⅤ࠾ࡼࡵ
573& ࡢࣗࢼ࣭ࢠெᕝᰶ㓗࡚ࠉ㧏ᗐᶭ⬗ࢅⓆ⌟ࡊࠉ
4'
3'
H
O
HO
ᵕࠍᚺ⏕◂✪ᙽࡈࡿࡾࠊ
1'
2'
B
ୌ⯙࡞ࣅ࣐ࣛࢩࣤࢽࢠ࢛ࣝࢨࢺࡡሲᇱ㒂ࡢ⁈ᾦ୯࡚ࠉ
5
6
6
5
H
4
3 ppm
4
3 ppm
H1'
H
NOE
H
H5
O
OH
7
6
5
1
Fig. 7 H NMR and NOE differential spectra of
5’-NH2-Urd obtained with presaturation at H6
(δ7.6) in (a) phosphate buffer (pH7.2) and (b)
borate buffer (pH 7.2).
ሲᇱヾㆉ࡞ฺ᭯ FQWL 㒼ྡྷࢅඁࡌࡾࠊࣛ࣍ࢽࢠ࢛ࣝ
ࢨ ࢺ ⢶ 㒂 Ề 㓗 ᇱ ࡞ ᯣ ᶣ ᵋ ㏸ ࢅ ᭯ ࡌ ࡾ
H:25 㸝 XULILQH ’’H^HOLH PRQRSKRVSKFWH 㸞 ࡷ
.5:㸝
4LVRSURS^OLIHQHXULILQH㸞
㒂ࡡ❟మ㝸ᐐ΅ࡊࡒ⤎ᯕ V^Q FQWL Ẓቌຊ
ࡊ࡙࠷ࡾࡆ࡛ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࡆࡡࡻ࠹
ࢽࢠ࢛ࣝࢨࢺㄇᑙమࡡሔྙࠉභ᭯⤎ྙ࡞ࡻࡽ
V^Q 㒼 ྡྷ ඁ ࡈ ࡿ ࡾ ࡒ ࡴ ࠉ አ 㒂 ᅄ Ꮔ ࡞ ࡻ ࡽ
FQWLV^Q㒼ྡྷࡡྊ㏣Ⓩንࢅㄇ㉫ࡌࡾࡆ࡛ࡢ
ᅏ㞬࡚࠵ࡾࠊአ㒂ᅄᏄ࡞ࡻࡾ V^Q 㒼ྡྷࡡㄇ㉫ࢅ㐡
10.0
[! ] x 10 -3 /deg cm2 dmol-1
ࡡㄇᑙమ࡚ࡢࠉሲᇱ㒂 న࢜ࣜ࣍ࢼࣜᇱ࡛ࠉ⢶
Urd
in phosphate buffer
Urd
in borate buffer
5.0
5'-NH2-Urd
in borate buffer
0.0
5'-NH2-Urd
in borate buffer
ᠺࡌࡾࡒࡴࠉ573& ࡚ࡢࠉ㸦㸞ሲᇱ㒂 న࢜ࣜ࣍ࢼ
ࣜ㓗⣪࡛ࣆࣚࢿ࣭ࢪ㒂 నỀ⣪㛣ࡡỀ⣪⤎ྙᙟ
ᠺࠉ㸧㸞ࣆࣚࢿ࣭ࢪ㒂 HLVࢩ࢛࣭࡛ࣜ࣌
㓗㢦࡛ࡡᯣᶣᵋ㏸ᙟᠺࡡ༝ྜྷຝᯕࡡฺ⏕ࢅ⩻᱄ࡊ
ࡒ㸝ࢲ࣭ࣔࢹ 㸞
ࠊ࣌㓗㢦ࡢ HLVࢩ࢛࣭࡛ࣜ
- 5.0
210
250
300
350
Wavelength/nm
Fig. 8 CD spectra of Urd and 5’-NH2-Urd in phosphate and
borate buffer.
Ề୯࡚ྊ㏣Ⓩ࡞࢙ࢪࢷࣜࢅᙟᠺࡌࡾࡆ࡛▩ࡼࡿ
࡙࠷ࡾࠊࣛ࣍ࢽࢠ࢛ࣝࢨࢺ࡞࠽࠷࡙ࡵ న࡞ HLVࢩ࢛࣭ࣜࢅ᭯ࡊࠉ࣌㓗࢙ࢪࢷࣜᙟᠺ࡞ࡻࡾ⢶㒂
ࢤࣤࣆ࢚࣒࣭ࢨࣘࣤን࡞ఔ࠹ሲᇱ㒂ࡡ㒼ྡྷንᙽࡈࡿࡾࠊࡈࡼ࡞ࣛ࣍ࢽࢠ࢛ࣝࢨࢺࡡ నỀ㓗ᇱ
11
ࢅ࣐ࢿᇱ࡞ንᥦࡊࡒ ࣐ࢿ
ࢸ࢛࢞ࢨࣛ࣍ࢽࢠ࢛ࣝࢨࢺࢅヾㆉ㒂న࡛ࡊ࡙⏕࠷ࡾ࡞ࡻࡽࠉຝ⋙
ࡻ࠷⢶㒂̺ᰶ㓗ሲᇱ㛣Ề⣪⤎ྙᙟᠺࡵᙽࡈࡿࡾࠊࡆࡡふⅤ࠾ࡼ నỀ㓗ᇱࢅ࣐ࢿᇱ࡞ንᥦࡊࡒ ࣐ࢿ
ࢸ࢛࢞ࢨࣛࢩࣤ㸝
3-:UI㸞ࢅྙᠺࡊࠉሲᇱ㒂㒼ྡྷࢅ -32734* ࢪ࣋ࢠࢹࣜ㸝ᅒ㸦㸞
ࡼࡦ࡞ළⰅ()ࢪ࣋ࢠࢹࣜ㸝ᅒ㸬㸞࡞ࡻࡽ᳠ゞࡊࡒࠊ
ࣛࣤ㓗⥾⾢ᾦ୯࡚ࡡ 3-:UI ࡡࣅ࣐ࣛࢩࣤሲᇱ㒂 నỀ⣪ࡡ↯ᑏ࡞ࡻࡽࠉሲᇱ㒂 న࣬⢶㒂 నỀ
⣪࡞ຊ࠻࡙⢶㒂 ࡐࡊ࡙ నỀ⣪࡞ࡵᰶ࢛࣭ࣁ࣭ࣀࢧ࣭ຝᯕ㸝34*㸞࡞ᇱࡘࡂࣅ࣭ࢠふῼࡈࡿࠉ
ሲᇱ㒂࢜ࣜ࣍ࢼࣜ㒂న⢶㒂ࡡཬᑊࢅྡྷࡂ FQWL 㒼ྡྷࢅඁࡌࡾࡆ࡛♟ြࡈࡿࡒࠊୌ᪁ࠉ⢶㒂 ’’HLV
ࢩ࢛࣭ࣜࠉᯣᶣᵋ㏸ࢅ᭯ࡌࡾ⎌≟࣌㓗࢙ࢪࢷࣜࢅᙟᠺࡌࡾ࣌㓗Ꮛᅹୖ࡚ࡢࠉྜྷࡋࡂሲᇱ㒂 నỀ
⣪↯ᑏ࡞ࡻࡽሲᇱ㒂 నỀ⣪࣬⢶㒂 నỀ⣪࡞ࡡࡲ 34* ࣅ࣭ࢠふῼࡈࡿࠉࡡ⢶㒂Ề⣪࡛ࡡ 34* ࣅ࣭
ࢠࡢධ࡙ᾐࡊࡒࠊ௧୕ࡡᐁࡢࠉ࣌㓗ࡡ῟ຊ࡞ࡻࡽᰶ㓗ሲᇱ㒂 FQWL ࠾ࡼ V^Q 㒼ྡྷඁ࡞ንࡊࡒ
ࡆ࡛ࢅ᪺☔࡞♟ြࡊ࡙࠷ࡾࠊࡌࢂࡔࠉ’࣐ࢿࣅ࣐ࣛࢩࣤࣛ࣍ࢽࢠ࢛ࣝࢨࢺࡢ࣌㓗㢦ࢅአ㒂ᅄᏄ࡛
ࡊ࡙ FQWLЍV^Q ሲᇱ㒂㒼ྡྷโᚒྊ⬗ฦᏄ࡚࠵ࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊ
ࡈࡼ࡞ 3-:UI ࡡ () ࢪ࣋ࢠࢹࣜࢅ࣌㓗⥾⾢ᾦ୯࡚ῼᏽࡌࡾ࡛@ θBPF] ೋࡢ FQWL 㒼ྡྷࢅ♟ࡌ ࠾
ࡼࠉV^Q 㒼ྡྷ࡞ᇱࡘࡂ ࡞ࡱ࡚΅ᑛࡊࡒࠊࡌࢂࡔࠉ() ࢪ࣋ࢠࢹࣜ࠾ࡼࡵ࣌㓗㢦࡞ࡻࡾ 㛣ࡡ
ᯣᶣᵋ㏸ᙟᠺࠉࡼࡦ࡞ฦᏄහỀ⣪⤎ྙᙟᠺࡡ༝ྜྷຝᯕ࡞ࡻࡽ࣌㓗ࢅአ㒂ᅄᏄ࡛ࡌࡾ V^Q 㒼ྡྷࡡㄇ㉫
Table 1
Anti-syn orienta
H2N
[! ]ext
phosphate buffer (a)
borate buffer (b)
b/a (%)
HO
phosphate buffer (a)
[! ]ext
borate buffer (b)
b/a (%)
㐡ᠺࡈࡿࡾࡆ࡛᪺ࡼ࠾࡞ࡖࡒࠊࡱࡒࠉྜྷࡋࡂࣅ࣐ࣛࢩࣤሲᇱࢅ᭯ࡊ Ề㓗ᇱࢅ࣐ࢿᇱ࡞ንᥦࡊ
ࡒ ࣐ࢿ
ࢸ࢛࢞ࢨࢨࢲࢩࣤ࡞࠽࠷࡙ࡵྜྷᵕ࡞࣌㓗㢦ࢅአ㒂ᅄᏄ࡛ࡊ࡙ሲᇱ㒂㒼ྡྷࢅ FQWLс V^Q
࡞㒼ྡྷโᚒྊ⬗࡚࠵ࡾࡆ࡛♟ࡈࡿࡒࠊୌ᪁ࠉ
నỀ㓗ᇱᏋᅹࡊ࠷ ࣐ࢿࢲ࣐ࢩࣤࡷ న࢜ࣜ࣍
12
ࢼࣜᇱᏋᅹࡊ࠷ ࣐ࢿࣛ࣍ࢸࢿࢨࣤࡷ ࣐ࢿࢿࢨ࡚ࣤࡢ࣌㓗῟ຊ࡞ఔ࠹ሲᇱ㒂㒼ྡྷࡢ
ධࡂふῼࡈࡿࡍࠉ㟸ᖏ࡞ᵋ㏸≁␏ᛮࡡ㧏࠷㒼ྡྷโᚒἪ࡚࠵ࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊ⾪ ࡞ᵕࠍࢽࢠ
࢛ࣝࢨࢺㄇᑙమ࡞ᑊࡊ࡙࣌㓗㢦࡞ࡻࡾྊ㏣Ⓩሲᇱ㒂㒼ྡྷโᚒ࡞ࡗ࠷࡙᳠ゞࡊࡒ⤎ᯕࢅ♟ࡊࡒࠊ
HN
O
O
N
HN
N
N
N
H
H
H
O
O
O
O
HN
O
H
N
O
N
N
H
N
O
O
N
O
N
H
O
O
HO OH
HO OH
O
HO OH
O
"-PRNA
!-PRNA
Chart 7 Structures of PRNAs.
ࡆࡡ⤎ᯕࢅ㊻ࡱ࠻ࠉ
࣐ࢿࣛ࣍ࢽࢠ࢛ࣝࢨࢺࢅᑙථࡊࡒ᩺ࡊ࠷࢜ࢷࢥ࣭ࣛࡡெᕝᰶ㓗࡛ࡊ࡙ࠉ࣋ࣈ
ࢲࢺࣛ࣍ᰶ㓗573&ࢅスゝ࣬ྙᠺࡊࡒ㸝ࢲ࣭ࣔࢹ㸬㸞
ࠊெᕝᰶ㓗࡞࠽࠷࡙ࡵࠉ࣌㓗㢦ࢅአ㒂ᅄᏄ࡛ࡌࡾ
࣐ࢿࣅ࣐ࣛࢩࣤࣛ࣍ࢽࢠ࢛ࣝࢨࢺሲᇱ㒂㒼ྡྷࡡ FQWLсV^Q 㒼ྡྷโᚒࢅᐁ⌟ࡌࡾࡒࡴ࡞ᚪさᵋ㏸᮪
௲࡛ࡊ࡙ࠉ㸦㸞࣌㓗࡛ྊ㏣Ⓩ࡞࣌㓗࢙ࢪࢷࣜࢅᙟᠺࡌࡾࡒࡴ࡞ᚪさ HLVࢩ࢛࣭ࣜࠉ㸧㸞ሲ
ᇱ㒂 న࢜ࣜ࣍ࢼࣜᇱ࡛Ề⣪⤎ྙᙟᠺྊ⬗ నỀ⣪⤎ྙ౩ᇱࢅ᭯ࡌࡾᚪさ࠵ࡾࠊࡊ࠾ࡊࠉࡆࡡᵕ
ᵋ㏸Ⓩᚪさ᮪௲ࡢኮ↓ )3&࣬73& ࡚ࡢᙔ↓㊂ࡈࡿࡍࠉᚉᮮሒ࿈ࡈࡿ࡙࠷ࡾ 53& ࡵྱࡴࡒ⭶ኬ⛸㢦ࡡ
ᰶ㓗࣓ࢸࣜྙ∸࣬ெᕝᰶ㓗࡚ࡵ㊂ࡈࡿ࠷ࠊࡐࡆ࡚ࠉ㸦㸞࡛㸧㸞ࡡ᮪௲ࢅ㊂ࡌࡾ࡛භ࡞ࠉᵕࠍ
ሲᇱ㒼าࢅ᭯ࡌࡾ࣓ࢸࣜᐖ᪾࡞ྙᠺྊ⬗࡚࠵ࡽࠉኮ↓ᰶ㓗࡛ྜྷ⛤ᗐࡡᰶ㓗ሲᇱࡡ⧖ࡽ㏁ࡊ㊝㞫ࢅ᭯ࡌ
ࡾᰶ㓗࣓ࢸ࡛ࣜࡊ࡙Ț573& ࢅスゝ࣬ྙᠺࡊࡒࠊࡱࡒࠉ㙈࡛ࡊ࡙ȘKHOL]ᵋ㏸㧏ḗᵋ㏸ࢅ᭯ࡌࡾ
ࡆ࡛ᙽࡈࡿࡾȘ573& ࡵスゝ࣬ྙᠺࡊࡒ ࠊ 13
Ș573&ࠉȚ573& ࠷ࡍࡿࡡ࣓ࢿ࣏Ѹ࡞࠽࠷࡙ࡵࣛࣤ㓗⥾⾢ᾦ୯࡚ࡢࣅ࣐ࣛࢩࣤሲᇱ㒂ࡢ FQWL 㒼ྡྷࢅඁ
ࡌࡾࡡ࡞ᑊࡊࠉ࣌㓗῟ຊ࡞ఔ࠷ V^Q 㒼ྡྷඁ࡞ንࡌࡾࡆ࡛ ’࣐ࢿࢽࢠ࢛ࣝࢨࢺࡡሔྙ࡛ྜྷᵕ
࡞ -32734*() ࢪ࣋ࢠࢹࣜࢅ⏕࠷ࡒ᳠ゞ࡞ࡻࡽ᪺ࡼ࠾࡛ࡖࡒࠊࡈࡼ࡞ሲᇱ㒂㒼ྡྷࡡྊ㏣Ⓩโᚒ࡞ࡗ
࠷࡙᳠ゞࡊࡒࠊ࣌㓗࢙ࢪࢷࣜࡡᏭᏽᛮ S- ࡞ኬࡀࡂ౪Ꮛࡊࠉࣜ࢜ࣛᛮ᮪௲ୖ࡚ࡢᏭᏽ࡞Ꮛᅹࡌࡾ࣌
㓗࢙ࢪࢷࣜࡢࠉ㓗ᛮ᮪௲ୖ࡚ኬࡀࡂᏭᏽᛮୖࡊࠉよ㞫᪁ྡྷ࡞ᖲ⾦⛛ິࡌࡾࡆ࡛࡞╌┘ࡊࡒࠊ࣌
㓗㢦ࢅ㒼ྡྷぞโᅄᏄࠉS- ࢅ㒼ྡྷโᚒᅄᏄ࡛ࡌࡾ ࣐ࢿࣅ࣐ࣛࢩࣤࢽࢠ࢛ࣝࢨࢺࡡྊ㏣Ⓩሲᇱ㒂
㒼ྡྷโᚒ࣌㓗࢙ࢪࢷࣜᙟᠺࡢ⣌ࡡ S- ࡞㟸ᖏ࡞ᨼវ࡚࠵ࡽࠉࣜ࢜ࣛᛮ᮪௲ୖ࡚ࡢᏭᏽ࡞Ꮛᅹࡌࡾࡡ࡞
pH 6.0
pH 8.8
[ ! ] x 10-3/deg cm2 dmol-1
ᑊࡊࠉ㓗ᛮ᮪௲ୖ࡚ࡢよ㞫ࡌࡾࡆ࡛▩ࡼࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉ࣌▹㸝3F
S-7]/mM
ࢅአ㒂ᅄᏄ࡛
'4㸞Ꮛᅹୖ
[Na2B4 O 7]/mM
[Na2B4 O
5
0
0
ࡊ࡙ 573& ༟㔖మࡡሲᇱ㒂ࡡྊ㏣Ⓩ㒼ྡྷโᚒ࡞ࡗ࠷࡙᳠ゞࡊࡒࠊࡱࡍࠉ࠷ࡂࡗ࠾ࡡ
S- ࡞࠽ࡄࡾ࣌▹῟
15
0.25
30
0.5
ຊ࡞ఔ࠹ () ࢪ࣋ࢠࢹࣜን࡞ࡗ࠷࡙᳠ゞࡊࡒ㸝ᅒ㸨㸞
ࠊ࣌㓗࢙ࢪࢷࣜᏭᏽ࡞Ꮛᅹࡌࡾ 0.75
S- ࡡᘽ
1.5
0
ࣜ࢜ࣛᛮ᮪௲ୖ࡚ࡢᑛ㔖ࡡ࣌▹῟ຊ࡞ఔ࠷
QP ㎾ࡡ () ᙁᗐࡢኬࡀࡂ΅ᑛࡊࠉຝ⋙Ⰳࡂሲᇱ㒂ࡡ
2.5
6.5
FQWLЍV^Q 㒼ྡྷโᚒ♟ࡈࡿࡒࡡ࡞ᑊࡊࠉ࣌㓗࢙ࢪࢷࣜࡡᏭᏽᛮୖࡌࡾ S- ࡡᘽ㓗ᛮ᮪௲ୖ࡚
ࡢኬ㔖ࡡ࣌
▹ࢅ῟ຊࡊ࡙
ࡵ () ࢪ࣋ࢠࢹ
-5
Kb = 156 M-1
- 10
[uridine unit] = 1.1 x 10-4 M
ࣜࡢ࡛ࢆ
ን ࡎ ࡍ ࠉ
’’ ࣌ 㓗
࢙ࢪࢷࣜᙟᠺ
ࢅ㘵 ࡛ ࡌ ࡾ
210
250
300
Wavelength/nm
350 210
[uridine unit] = 1.1 x 10-4 M
250
300
350
Wavelength/nm
Figure 3. CD spectral change of PRNA monomer containing uracil base by adding borax to
phosphate buffer at pH 6.0 (left) and pH 8.8 (right).
ㄇ㉫ࡈࡿ࠷ࡆ࡛᪺ࡼ࠾࡛ࡖࡒᅒ㸨ࠊࡆࡡ⤎ᯕࢅࡨࡱ࠻ࠉ࣌▹ᏋᅹୖࠉS- ࢅአ㒂ᅄᏄ࡛ࡊࡒሲᇱ㒂㒼ྡྷ
᳠ゞࡊࡒᅒ㸩ࠊS- ࡡࣛࣤ㓗⥾⾢⁈ᾦ࡞ P2 ࡡ࣌▹ࢅ῟ຊࡌࡾ࡛@θBH]W ࡢ΅ᑛࡊሲᇱ㒂ࡢ FQWLЍV^Q ࡞
⤾࠷࡙ࠉࡆࡡ⣌ࡡ S- ࢅ ࡞ㄢᩒࡌࡾ࡛@ θBH]W ࡢ࣌▹῟ຊ๑ࡡೋࡱ࡚ᅂᚗࡊࠉሲᇱ㒂ࡢ V^QЍFQWL 㒼ྡྷ࡞ንࡊ
ࢅ ⤾࠷࡙ ࡛ㄢᩒࡌࡾࡆ࡛࡞ࡻࡽࠉ@θBH]W ࡵྊ㏣Ⓩ࡞ንࡊࠉሲᇱ㒂㒼ྡྷ࡞ࡗ࠷࡙࣌▹Ꮛᅹୖ S- ࢅአ㒂
ᚒྊ⬗࡚࠵ࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕࡻࡽࠉ㒼ྡྷንࢅฺ⏕ࡊࡒᰶ㓗ヾㆉࡡྊ㏣Ⓩโᚒᙽࡈࡿࡾࠊ
14
௧୕ࡡ⤎ᯕࢅ㊻ࡱ࠻ࠉࣤࢲࢬࣤࢪฦᏄࡷ⏍మ㐲
ྙᛮᮞᩩࡡ㐲⏕࡞ྊḖ 573& ࢛ࣛࢥ࣏࣭
H2 N
RESIN
ྙᠺ࡞ཱིࡽ⤄ࢆࡓࠊࣤࢲࢬࣤࢪ࣬ࣤࢲࢩѸࣤฦ
Ꮔࡢࠉࢰ࣭ࢣࢴࢹ 73&࣬)3& ࡛┞Ⓩ⛸ࠍࡡ㒼า
NMP
20% PPD /NMP
ࢅ᭯ࡌࡾࡆ࡛ᚪさྊḖ࡚࠵ࡾࠊᠻࠍࡢᙔิࠉ'RH
Fmoc-ly s(Boc)-OH or Fmoc -Gln(5'N)-OH
Bop reagent, H OBt, NMP, DIEA
ᇱࢅ 3 ᮆ❻ಕ㆜ᇱ࡛ࡊ࡙⏕࠷ࡾᾦ┞Ἢ࡞ࡻࡽ 573&
࢛ࣛࢥ࣏࣭ྙᠺࢅ᳠ゞࡊ࡙ࡀࡒࠉᮇྙᠺἪࡡࣈࣛ
ࣤሲᇱࡡᣉᘿࢅ⩻࠻ࡾሔྙࠉ'RH ᇱࡡ⬲ಕ㆜㐛⛤
if negative
if pos itive
Kaise r test
࡞࠽ࡄࡾ㓗ᛮ᮪௲ୖ࡚ࠉࢽࢠ࢛ࣝࢨࢺ㒂ࡻࡽ⬲ࣈࣛ
20% PPD/NMP
28% NH 3aq
ࣤཬᚺࡡ㐅⾔ࡵࡈࡿࡾࠊࡈࡼ࡞ࡢᵕࠍ㒼า
ࢅ᭯ࡌࡾ 573& ࢛ࣛࢥ࣏࣭ࡡ⠾౼࠾ࡗ㧏⋙ྙᠺ
2.5% H 2O/TFA, 2.5% TIS
precipitated with Et2 O
R. P. HPLC purification
ᅏ㞬࡚࠵ࡾ࡛࠷ࡖࡒᮇ㈻Ⓩၡ㢗Ⅴࡵ࠵ࡖࡒࠊୌ
᪁ࠉIOXRUHQ^OPHWK^OR]^HFUGRQ^O+PRHᇱࢅ 3
ᮆ❻ಕ㆜ᇱ࡛ࡊ࡙⏕࠷ࡾ࣋ࣈࢲࢺᅖ┞ྙᠺἪ࡞ࡻ
ࡾα࣋ࣈࢲࢺྙᠺᩐኣࡂ᳠ゞࡈࡿ࡙࠷ࡾࠊ+PRH
Ἢࡢ ࣅ࣋ࣛࢩࣤ55)ࢅ⏕࠷ࡾῺᘽሲᇱ
PRNA oligomer
Figure 5. Solid-phase Peptide Synthesis using Fmoc Protective
Group
ᛮ᮪௲ୖ࡚ +PRH ᇱࢅ⬲ಕ㆜ࡌࡾࡒࡴࠉ573& ࡡࣈ
ࣛࣤࢽࢠ࢛ࣝࢨࢺ㒂⬲ࣈࣛࣤ࡞ᇱࡘࡂ࣭࣊ࢨ
Ὡᛮࢅ᭯ࡌࡾ⦨ྙࢅ᳠ゞࡌࡾࡆ࡛࡞ࡻࡽࠉ
573& ࡡࢽࢠ࢛ࣝࢨࢺ㒂ࡡ ’’Ề㓗ᇱࢅ↋ಕ㆜࡚
༎ฦఘ㛏ཬᚺຝ⋙ࡡ㐡ᠺྊ⬗࡞ࡾ࡛⩻࠻ࡼ
ࡿࡾࠊࡐࡆ࡚ඔᮮα࣐ࢿ㓗ࡡఘ㛏ཬᚺ࡞⏕࠷ࡼ
ࡿ࡙ࡀࡒ +PRH ᅖ┞ྙᠺἪࢅ⏕࠷ࡒ 573& ྙᠺ࡞ࡗ
࠷࡙᳠ゞࡊࡒࠊࡐࡡ⤎ᯕࠉ⮤ິ࣬༖⮤ິᅖ┞ྙᠺ
[! ] x 10-3/deg cm2 dmol-1
ࢴࢠࢦࢹ⏍ᠺࡡᢒโᙽࡈࡿࡾࠊࡈࡼ࡞ࠉᵕࠍ
6.0
[Na2B4 O7 ]/mM
0
0.06
0.12
0.20
0.40
0.79
1.36
1.92
3.70
3.0
0.0
- 3.0
Ἢࡡ㐲⏕ྊ⬗࡛ࡽࠉ⛸ࠍࡡ㒼าࢅ᭯ࡌࡾ 㔖మ⛤ᗐࡡ࢛ࣛࢥ 573& ࡵ⠾౼࠾ࡗ㧏⋙࡚ྙᠺ
ྊ⬗࡛ࡖࡒ㸝ᅒ㸪㸞ࠊ
- 6.0
210
250
ࡆࡡࡻ࠹࡞ࡊ࡙ྙᠺࡊࡒ࢛ࣛࢥ 573& ࡡ࣌㓗
㢦ࢅአ㒂ᅄᏄ࡛ࡌࡾ FQWLV^Q㒼ྡྷโᚒ࡞ࡗ࠷࡙
()32734* ࢅ⏕࠷࡙᳠ゞࡊࡒࠊᅒ㸫࡞࣓࣌ࣚ
300
350
Wavelength/nm
Figure 6. CD spectra changes of (isoGln(5'U))8-Lys･HCl with
increasing concentrations of Na2B4O7 in phosphate buffer; [uracil
base] = 1.0 x 10-4 M.
ࢨࣜ㒼าࢅ᭯ࡌࡾȚ573& 㔖మࡡࣛࣤ㓗⥾⾢ᾦ
࡞࣌▹ࢅ῟ຊࡊࡒ⣌࡞࠽ࡄࡾ () ࢪ࣋ࢠࢹࣜንࢅ♟ࡊࡒࠊࣛࣤ㓗⥾⾢ᾦ୯࡚ࡢ ⛤ᗐࡡ@ θBࢅ♟ࡊ
FQWL 㒼ྡྷࢅඁࡌࡾࡆ࡛♟ြࡈࡿࡒࡡ࡞ᑊࡊࠉ࣌▹ࢅ῟ຊࡌࡾࡆ࡛࡞ࡻࡽ 573& ࣓ࢿ࣏Ѹࡷ ࣐
ࢿࣅ࣐ࣛࢩࣤࣛ࣍ࢽࢠ࢛ࣝࢨࢺࡻࡽຝ⋙ࡻࡂ V^Q 㒼ྡྷㄇ㉫ࡈࡿࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊࡆࡡአ㒂ᅄ
Ꮔ࡞ࡻࡾሲᇱ㒂㒼ྡྷንࡢ -32734* ࢪ࣋ࢠࢹࣜ࡞ࡻࡖ࡙ࡵ☔ヾࡈࡿࠉ573& ࡢአ㒂ᅄᏄ࡞ࡻࡖ࡙ྊ㏣Ⓩ
࡞ሲᇱ㒂㒼ྡྷโᚒྊ⬗ࡢࡋࡴ࡙ࡡெᕝᰶ㓗࡚࠵ࡾࡆ࡛♟ࡈࡿࡒࠊ
15
᭩ᚃ࡞᭩ࡵ㔔さࢰ࣭ࢣࢴࢹ )3&࣬73& ࡛ࡡ㘊ᙟᠺᣪິࠉ㘊మᏭᏽᛮࠉࡐࡊ࡙㘊ᙟᠺ࣬よ㞫โᚒ⬗ࢅ᳠
ゞࡌࡾࡒࡴ࡞┞Ⓩሲᇱࢅ᭯ࡌࡾ࣎ࣛࢽࢠ࢛ࣝࢲࢺࢅ⏕࠷࡙ࠉ࣌㓗㢦῟ຊ࡞ఔ࠹㘊ᙟᠺ࣬よ㞫ᣪິ࡞
ࡗ࠷࡙᳠ゞࡊࡒࠊ⛸ࠍࡡ㒼าࢅ᭯ࡌࡾ 573& ࠽ࡻࡦ 㔖మ࢛ࣛࢥ࣏࣭ࡢࠉ+PRH ᅖ┞ྙᠺἪࢅ⏕࠷࡙ᚋ
ࡒࠊ573& ࢛ࣛࢥ࣏࣭ࡡሲᇱヾㆉ⬗ࡢ⼝よῺᗐ㸝9P㸞ࢅᣞᵾ࡛ࡊ࡙ࠉ┞Ⓩሲᇱࡼࡦ࡞࣐ࢪ࣏ࢴࢲ㒼า
ࢅ᭯ࡌࡾ )3& ࢅ⏕࠷࡙᳠ゞࡊࡒࠊࢤࣤࣈࣝࢴࢠࢪࡡሲᇱࣗࢼࢴࢹẒࡢ :; ࢪ࣋ࢠࢹࣜࡡᰶ㓗ሲᇱ㒂ࡡྺ
࡞ᇱࡘࡂ QP ࡞࠽ࡄࡾྺකᗐࡡ /RG ࣈࣞࢴࢹ࠾ࡼ 㸯 ࡚࠵ࡾࡆ࡛᪺ࡼ࠾࡛ࡽࠉ53& ࡢ┞㙈
)3& ࡛ ࡡ㘊మࢅᙟᠺࡌࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊࡱࡒࠉ㟸┞㙈࡛ࡡΊྙ⁈ᾦ࡚ࡢῈⰅຝᯕࠉ9P ࡛
ࡵ࡞ふῼࡈࡿ࠾ࡖࡒࡆ࡛࠾ࡼࠉ573& ࡛ )3& ࡡ㘊మࡢᰶ㓗ሲᇱ㛣ࡡ≁␏ⓏỀ⣪⤎ྙ࡞ࡻࡽᙟᠺࡈࡿ࡙࠷
ࡾࡆ࡛᪺ࡼ࠾࡞ࡖࡒࠊࣛࢩࣤࢅ᭯ࡌࡾ 573& 㔖మ࡛ I& ࡡࣛࣤ㓗⥾⾢ᾦ୯࡞࠽ࡄࡾ 9P ೋࡢࠉ)3&
㘊మࡡ 9P ࡛Ẓ㍉ࡌࡾ࡛ Υ㧏࠷ೋࢅ♟ࡊࠉ573&ࡢኮ↓ᰶ㓗࡛Ẓ㍉ࡊ࡙ࡻࡽᏭᏽ㘊మࢅᙟᠺࡌࡾࡆ࡛
᪺ࡼ࠾࡞ࡖࡒࠊࡱࡒࠉࣚࢨࣜ࠽ࡻࡦࢨࢹࢨࣤࢅஹப࡞᭯ࡌࡾ 573& 㔖మࡡሔྙࠉ࣐ࢿᮆ❻࡛ኮ↓
ᰶ㓗ࡡ ᮆ❻ࢅྜྷࡋഁ࡞᭯ࡌࡾ FQWLSFUFOHOOHᆵ㔔㙈ࠉ㏣ࡡ SFUFOOHOHᆵ㘊మࡻࡽᏭᏽ࡞Ꮛᅹࡌࡾ
ࡆ࡛♟ࡈࡿࠉ)3& 㙈᪁ྡྷ࡞ᑊࡌࡾⰃይㆉื⬗ࢅ᭯ࡌࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊ53& ࡞࠽࠷࡙ࡢ࡞㏑
ࡒࡻ࠹࡞ࠉ)3& 㙈᪁ྡྷ࡞ᑊࡌࡾヾㆉᛮ࠷ࡆ࡛࠾ࡼࠉࢽࢠ࢛ࣝࢨࢺࡡᑙථ࡛࣐࢞ࣚࣜࢺ㦭࡞ࡻࡽ
573& ࡢⰃይ㙈᪁ྡྷヾㆉᛮࢅ⋋ᚋࡊࡒ࡛⩻࠻ࡼࡿࡾࠊ
ࡆࡡࡻ࠹࡞ )3&࣬)3& 㘊మࡻࡽᏭᏽ 573&࣬)3& 㘊మ࡚࠵ࡾࠉ⁈ᾦ࡞ᑛ㔖ࡡ࣌㓗ࢅ῟ຊࡌࡾ࡛ )3&࣬
)3& 㘊మᆵ࡚ࡢሲຝᯕ࡞ࡻࡽ 9P ୕᪴ࡊࡒࡡ࡛ࡢᑊ↯Ⓩ࡞ࠉ࠷ࡍࡿࡡ 573&࣬)3& 㘊మᙟᠺ⣌࡚ࡵ 9P ࠽ࡻࡦ
ῈⰅຝᯕࡢふῼࡈࡿࡂࡽࠉᏭᏽ 573&࣬)3& 㘊మ㎷㏷࡞よ㞫ࡊࡒࡆ࡛♟ࡈࡿࡒࠊࡆࡡࡻ࠹ຝ
ᯕⓏ㘊よ㞫ࡢࠉ573& ࡡሲᇱ㒂࣌㓗῟ຊ࡞ࡻࡽ V^Q 㒼ྡྷࢅඁࡌࡾࡒࡴ )3& ࡛ࡡሲᇱᑊᙟᠺᅏ㞬
࡞ࡖࡒࡆ࡛࡛ࠉ࣌㓗࢙ࢪࢷࣜᙟᠺ࡞࡛ࡵ࠹࣌⣪୕࡞⏍ࡋࡾㇿ㞹Ⲭ࡞ࡻࡾ )3& 㙈࡛ࡡ㟴㞹Ⓩཬ
Ⓠࡡ༝ྜྷຝᯕ࡞ᇱࡘࡂ࡛⩻࠻࡙࠷ࡾࠊࢲ࣭ࣔࢹ㸭
ࡆࡡࡻ࠹࡞ 573& ࡢ┞Ⓩ )3& ࡛ሲᇱ㒼า≁␏Ⓩ࡞Ꮽᏽࢤࣤࣈࣝࢴࢠࢪࢅᙟᠺࡌࡾࡆ࡛᪺ࡼ࠾࡛
ࡖࡒࡡ࡚ࠉአ㒂ᅄᏄ࡞ࡻࡾྊ㏣Ⓩࢤࣤࣈࣝࢴࢠࢪᙟᠺ࣬よ㞫โᚒ࡞ཱིࡽ⤄ࢆࡓࠊරమⓏ࡞ࡢࡡ
ࢽࢠ࢛ࣝࢨࢺㄇᑙమࡼࡦ࡞ 573& ࣓ࢿ࣏Ѹ⣌࡞᪂࠷࡙࣌▹ᏋᅹୖࠉS-ࢅአ㒂ᅄᏄ࡛ࡌࡾྊ㏣Ⓩ FQWL
сV^Q 㒼ྡྷโᚒࡡ⤎ᯕࢅ㊻ࡱ࠻ࠉ573&)3& ࢤࣤࣈࣝࢴࢠࢪᙟᠺ⣌࡞࣌▹ࢅ῟ຊࡊࠉS-ࢅአ㒂ᅄᏄ࡛ࡊ
ࡒࢤࣤࣈࣝࢴࢠࢪᙟᠺ࣬よ㞫ᣪິࢅ :; ࢪ࣋ࢠࢹࣜ༟Ⰵຝᯕࡼࡦ࡞ 9P ࢅᣞᵾ࡛ࡊ࡙᳠ゞࡊࡒࠊ573&)3&
ࢤࣤࣈࣝࢴࢠࢪᙟᠺ⣌࡞࣌▹ࢅ῟ຊࡌࡾࡆ࡛࡞ࡻࡽᏭᏽ࡞ᙟᠺࡈࡿ࡙࠷ࡒ 573&)3& 㘊మࡢ㎷㏷࠾ࡗ㧏
ຝ⋙࡞よ㞫ࡊࠉࢤࣤࣈࣝࢴࢠࢪᙟᠺࡡ࢛ࣤЍ࢛ࣆโᚒ㐡ᠺࡈࡿࡒࠊࡗࡁ࡞࣌㓗῟ຊ⣌ࡡ S- ࢅ ࠾
ࡼ ࡞ㄢᩒࡌࡾࡆ࡛࡞ࡻࡽῈⰅຝᯕࡢ්ࡦᅂᚗࡊ࣌▹῟ຊ๑࡛ྜྷࡋ 9P ふῼࡈࡿࠉሲᇱ㒂ࡡ V^QЍ
FQWL 㒼ྡྷඁ࡞ఔ࠷ࠉ573&)3& 㘊మࡡ්ᙟᠺ☔ヾࡈࡿࡒࠊࡌࢂࡔ 573& ࡢ࣌▹ᏋᅹୖࠉS- ࢅአ㒂
ᅄᏄ࡛ࡊ )3& ヾㆉࡡྊ㏣Ⓩโᚒᶭ⬗ࢅ᭯ࡌࡾࡆ࡛᪺ࡼ࠾࡛ࡖࡒࠊ
ࡆࡡࡻ࠹࡞ 573& ࡢ⣵⬂㈻හ S- ን࡞ᑊᚺࡌࡾࢂࡍ࠾ S-ࣝࣤࢩ࡚ࢰ࣭ࢣࢴࢹ )3&࣬73& ࡞ᑊࡌࡾヾ
ㆉ࣬㘊మᙟᠺࡡ࢛ࣤс࢛ࣆโᚒᶭ⬗ࢅ᭯ࡌࡾࡆ࡛♟ࡈࡿࡒࠊ᭩㎾࢝ࣤ⣵⬂ࡡ⣵⬂㈻ S-ࡢࢨࣜ㓗
ࡡ㓗ᛮኣ⢶ࡡ㐛Ⓠ⌟࡞ࡻࡽḿᖏ⣵⬂ࡡ⣵⬂㈻ S- ࡻࡽୖࡊࠉࠤ ࡚࠵ࡾࡆ࡛ሒ࿈ࡈࡿ
ࡒࠊࡆࡡሒ࿈࡞ᇱࡘࡂ࡛ࠉ573&࣌㓗⤎ྙమࡢࠉḿᖏ⣵⬂හ࡚ࡢ V^Q 㒼ྡྷࢅඁࡊࠉࣤࢲࢬࣤࢪᶭ⬗
ࢅⓆ⌟ࡊ࠷ࡡ࡞ᑊࡊࠉ࢝ࣤ⣵⬂ࡡ㓗ᛮ⣵⬂㈻࡞࠽࠷࡙ࡢ 573&࣌㓗⤎ྙమよ㞫ࠉࣤࢲࢬࣤࢪᶭ⬗
ࢅⓆ⌟ࡌࡾࡆ࡛ᙽ࡚ࡀࡾࠊࡆࡡࡆ࡛ࡢ≁␏ᛮⱕᖱୖࡌࡾࡵࡡࡡࠉ㧏࠷Ề⁈ᛮࡷ㧏ḗᵋ㏸ࢅᣚࡒ
16
࠷ࣤࢲࢬࣤࢪฦᏄ࡛ࡊ࡙ඁࡿࡒ≁ᛮࢅ᭯ࡌࡾ ࠤ 㔖మ⛤ᗐࡡ࢛ࣛࢥெᕝᰶ㓗ࢅࠉ⌟ᅹᙽ᭻ࡈ
ࡿ࡙࠷ࡾ࢝ࣤ⣵⬂≁␏ⓏࣤࢲࢬࣤࢪฦᏄࡡᒈ㛜ࢅྊ⬗࡛ࡌࡾ᪁Ἢㄵ࡚࠵ࡾ࡛ᙽࡈࡿࠉ⌟ᅹ⣵⬂ࣝ
࡚࣊ࣜࡡ◂✪ࢅゝ⏤୯࡚࠵ࡾࠊ
௧୕ࡡࡻ࠹࡞ 3LHOVHQ ࡼ࡞ࡻࡽᥞ᱄ࡈࡿඁࡿࡒᛮ㈻ࢅ♟ࡌ࣋ࣈࢲࢺᰶ㓗㸝53&㸞ࡡ᩺ࡊ࠷ᒈ㛜࡛ࡊ࡙ࠉ
ᠻࠍࡢ࣋ࣈࢲࢺࣛ࣍ᰶ㓗㸝573&㸞࡛࠷࠹᩺ࡊ࠷࢜ࢷࢥ࣭ࣛࡡฦᏄࢅ⏕࠷ࡾࡆ࡛࡞ࡻࡽࠉᚉᮮࡡ༟ࡾᰶ
㓗ࡡヾㆉ࠾ࡼୌṄ㐅ࢆ࡚አ㒂ᅄᏄ࡞ࡻࡾᰶ㓗ヾㆉโᚒࢅ⾔࠹᩺ࡒ᪁Ἢㄵࢅᥞ᱄ࡊࠉ࠷ࡂࡗ࠾ࡡරమౚ
࡚ࡐࡡ᭯ຝᛮࢅᐁッࡊࡒࠊࡆࡡ᪁Ἢㄵࡢୌ⯙ᛮࢅ᭯ࡊࠉ⥬ゕ࡚㏑ࡒḗୠࡡ㐿ఎᏄ⒢⏕ࣤࢲࢬࣤ
ࢪฦᏄࡡࡲࡼࡍࠉ)3& ࢲࢴࣈࡡ㐿ఎᏄチ᩷ⷾࡷฦᏄ⏍∸Ꮥࡡᚺ⏕ࡵྊ⬗࡚࠵ࡽࠉᚃࡈࡼࡾⓆ
ᒈᙽࡈࡿࡾࠊ
H NH
O
O
H
N
HN
O H NH
N
N
O
O
O
N
N H N
O
N
OHOH
Complexation
O
O
N
Borax
HN
P
O
O
O
H
HN
O
O
N
O
N
O
N
N
N
+
O O
B
HO
OH
O
O
P
O
O
Decomplexation
Chart 8 The schematic drawing of reversible complexation and decomplexation processes of
PRNA-DNA mixing system by Na2B4O7 as external factor.
17
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a) H. Asanuma, M. Komiyama, Angew. Chem. Int. Ed. 1999, 38, 2393; b) H. Asanuma, M. Komiyama,
Angew. Chem. Int. Ed. 2001, 40, 2671; c) H. Asanuma, M. Komiyama, J. Am. Chem. Soc. 2002, 124, 1977.
28)
T. Wada, N. Minamimoto, Y. Inaki, Y. Inoue, Chem Lett, 1998, 1025.
a) T. Wada, N. Minamimoto, Y. Inaki, Y. Inoue, Nucleic Acids Res. Symp. Ser, 1998, 39, 29;. b) T. Wada,
N.
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19
Figure & Chart Captions
Chart 1 Structures of modified nucleotides.
Chart 2 DNA, PNA, and PNA derivatives.
Chart 3 Synthetic scheme of ε-Dde protected lysine.
Chart 4 Schematic drawing of molecular beacon PNA.
Chart 5
The schematic drawing of reversible control of recognition and complexation behavior of
PRNA by borates.
Chart 6
Strategy for active control of nucleobase orientation by borates.
Chart 7 Structures of PRNAs.
Chart 8 The schematic drawing of reversible complexation and decomplexation processes of
PRNA-DNA mixing system by Na2B4O7 as external factor.
Figure 1 1H NMR and NOE differential spectra of 5’-NH2-Urd obtained with presaturation at H6 (δ7.6)
in (a) phosphate buffer (pH7.2) and (b) borate buffer (pH 7.2).
Figure 2 CD spectra of Urd and 5’-NH2-Urd in phosphate and borate buffer.
Figure 3 CD spectra changes of PRNA monomer containing
uracil base by adding Na2B4O7 to
phosphate buffer at pH 6.0 (left) and pH 8.8 (right).
Figure 4 PH dependence of [θ
ext
of PRNA monomer containing uracil base in the presence of Na2B4O7.
Figure 5 Solid-phase peptide synthesis using Fmoc protective group.
Figure 6 CD spectra changes of (isoGln(5'U))8-Lys･HCl with increasing concentrations of Na2B4O7 in
phosphate buffer; [uracil base] = 1.0 x 10-4 M.
Table 1 Anti-syn orientation of nucleosides and nucleoside derivatives