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Supplementary Information for
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Efficient Cycloreversion of cis,syn-Thymine Photodimer by a Zn2+–Cyclen Complex
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Bearing a Lumiflavin and Tryptophan by Chemical Reduction and Photoreduction of a
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Lumiflavin Unit (Cyclen = 1,4,7,10-Tetraazacyclododecane).
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Yasuyuki Yamada and Shin Aoki
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Faculty of Pharmaceutical Sciences, Tokyo University of Science
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2641Yamazaki, Noda 278-8510, Japan
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((ZnL4)ox(H2O)), (b) 0.5 mM T[c,s]T (3), and (c) 0.5 mM 11ox ((ZnL4)ox(H2O)) + 0.5 mM T[c,s]T (3) in
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10% MeCN aqueous solution with I = 0.1 (NaNO3) at 25˚C.
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Page 5:
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T[c,s]T (3) in 10% MeCN aqueous solution with I = 0.1 (NaNO3) at 25˚C.
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Figure S1. Typical potentiometric pH titration curves for a solution of (a) 0.5 mM 11ox
Figure S2. Distribution diagram for a mixture of 100 M 11ox ((ZnL4)ox) and 400 M
Figure S3. Cyclic voltammogram of (a) 0.3 mM 10ox (solid line) and (b) 0.3 mM 11ox
28 (dashed line) on a glassy carbon disk in 0.1 M nBu4NClO4–CH3CN at a scan rate of 10 mV·s–1.
1
1 Page 7:
Figure S4. Change in UV-Vis absorption spectra (A) and fluorescence emission spectra
2 (B) of 3-carboxymethyllumiflavin 16 during photoirradiation in the presence of 35 mM Et3N in an
3 aqueous solution (pH 11) at 25°C: (a) spectra before UV irradiation, (b) after UV irradiation for 30 min,
4 and (c) after reoxidation in air. Change in UV-Vis absorption spectra (C) and fluorescence emission
5 spectra (D) of 16 during the photoirradiation in the presence of 35 mM DABCO in an aqueous solution
6 (pH 11) at 25°C: (d) spectra before UV irradiation, (e) after UV irradiation for 30 min, and (f) after
7 reoxidation in air.
8 Page 8:
[16] = 50 M.
Irradiation wavelength was 365 nm.
Figure S5. Change in UV-Vis absorption spectra (A) and fluorescence emission spectra
9 (B) of 3-carboxymethyllumiflavin 16 during photoirradiation in the presence of 35 mM potassium
10 oxalate in an aqueous solution (pH 11) at 25°C: (a) spectra before UV irradiation, (b) after UV
11 irradiation for 30 min, and (c) after reoxidation in air. Change in UV-Vis absorption spectra (C) and
12 fluorescence emission spectra (D) of 16 during the photoirradiation in the presence of 35 mM EDTA in
13 an aqueous solution (pH 11) at 25°C: (d) spectra before UV irradiation, (e) after UV irradiation for 30
14 min, and (f) after reoxidation in air. [16] = 50 M.
15 Page 9:
Irradiation wavelength was 365 nm.
Figure S6. Reaction rates for photoreversion of T[c,s]T (3) at 20°C by
16 3-carboxymethyllumiflavin 16 in an aqueous solution with 35 mM Et3N (closed circles), or 35 mM
17 potassium oxalate (open circles), or 35 mM EDTA (open triangles), or 35 mM DABCO (open squares)
18 in an aqueous solution at pH 11 (pH was adjusted with aqueous NaOH).
[T[c,s]T] = 400 M, [16] =
19 100 M.
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Figure S7. UV spectra of 10ox (a) before and (b) after UV irradiation for 10 min at pH
21 11 in the absence of Et3N at 25°C and UV spectra of 11ox (c) before and (d) after irradiation for 10 min
22 and (e) after reoxidation in air.
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[10ox] = [11ox] = 50 M.
Figure S8. Change in UV-Vis absorption spectra of 10ox upon addition of 1 mM EDTA
24 in an aqueous solution (pH 11) at 25°C: spectra of (a) 10ox and (b) 10ox with 1 mM EDTA.
[10ox] = 50
25 M.
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Figure S9.
Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 (open
27 squares) and 11 (closed squares) in an aqueous solution (pH 11) with 35 mM DABCO in comparison
2
1 with that by 10 + Et3N (closed circle).
2 Page 13:
Figure S10.
Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 in an
3 aqueous solution with 35 mM EDTA at pH 7.6 (closed circles), or 35 mM EDTA at pH 11 (open circles),
4 or 1 mM EDTA at pH 11 (open triangles), or 35 mM Et3N at pH 11 (open squares).
5 Page 14:
Figure S11.
Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 in an
6 aqueous solution with 35 mM potassium oxalate at pH 7.6 (closed circles), or 35 mM potassium oxalate
7 at pH 11 (open circles), or 35 mM Et3N at pH 11 (open squares).
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10
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16
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3
1
Figure S1.
Typical potentiometric pH titration curves for a solution of (a) 0.5 mM 11ox
2
((ZnL4)ox(H2O)), (b) 0.5 mM T[c,s]T (3), and (c) 0.5 mM 11ox ((ZnL4)ox(H2O)) + 0.5 mM T[c,s]T (3) in
3
10% MeCN aqueous solution with I = 0.1 (NaNO3) at 25˚C.
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12
(b)
(a)
11
pH
10
9
8
(c)
7
6
0
7
0.5
1.0
1.5
–
equiv (HO )
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10
11
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16
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4
2.0
1 Figure S2.
Distribution diagram for a mixture of 100 M 11ox ((ZnL4)ox) and 400 M T[c,s]T (3) in
2 10% MeCN aqueous solution with I = 0.1 (NaNO3) at 25˚C.
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5
Relative concentration / %
100
(T[c,s]T)–
80
T[c,s]T
60
(ZnL4)ox – (T[c,s]T)– complex
(ZnL4)ox(HO–)
(ZnL4)ox(H2O)
40
20
0
6
6
7
9
8
pH
7
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9
10
11
12
13
14
15
16
17
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5
10
11
1 Figure S3. Cyclic voltammogram of (a) 0.3 mM 10ox (solid line) and (b) 0.3 mM 11ox (dashed line)
2 on a glassy carbon disk in 0.1 M nBu4NClO4–CH3CN at a scan rate of 10 mV·s–1.
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4
5
O
Me
N
N
O
N
N
2e- + 2H+
Me
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9
N
H
N
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4
10
R5
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O2
O
10ox or 11ox
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R5
1 2
N
H
N
H
O
N
O
H
(b)
10 A
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7
8
6
N
H N
(a)
– 0.4
E / V vs. Ag/Ag+
O
HN
3 R5
10red or 11red
– 0.8
R3
=
0
N
N
H
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Figure S4.
Change in UV-Vis absorption spectra (A) and fluorescence emission spectra (B) of
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3-carboxymethyllumiflavin 16 during photoirradiation in the presence of 35 mM Et3N in an aqueous
3
solution (pH 11) at 25°C: (a) spectra before UV irradiation, (b) after UV irradiation for 30 min, and
4
(c) after reoxidation in air. Change in UV-Vis absorption spectra (C) and fluorescence emission
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spectra (D) of 16 during the photoirradiation in the presence of 35 mM DABCO in an aqueous
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solution (pH 11) at 25°C: (d) spectra before UV irradiation, (e) after UV irradiation for 30 min, and (f)
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after reoxidation in air.
[16] = 50 M.
Irradiation wavelength was 365 nm.
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(A)
(B)
Fluorescence Intensity / a.u.
/ M–1 ·cm–1
 10–4
Et3N
3
2
(c)
(a)
1
(b)
250 300
400
Wavelength / nm
500
30
550
600
Wavelength / nm
650
(D)
150
Fluorescence Intensity / a.u.
/ M–1 ·cm–1
(b)
60
475 500
4
 10–4
90
600
(C)
3
(d), (e), (f)
2
1
(f)
120
(d)
90
60
(e)
30
0
0
250 300
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(a)
120
0
0
DABCO
(c)
150
4
400
500
Wavelength / nm
600
7
475 500
550
600
Wavelength / nm
650
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Figure S5.
Change in UV-Vis absorption spectra (A) and fluorescence emission spectra (B) of
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3-carboxymethyllumiflavin 16 during photoirradiation in the presence of 35 mM potassium oxalate in
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an aqueous solution (pH 11) at 25°C: (a) spectra before UV irradiation, (b) after UV irradiation for 30
4
min, and (c) after reoxidation in air. Change in UV-Vis absorption spectra (C) and fluorescence
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emission spectra (D) of 16 during the photoirradiation in the presence of 35 mM EDTA in an aqueous
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solution (pH 11) at 25°C: (d) spectra before UV irradiation, (e) after UV irradiation for 30 min, and (f)
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after reoxidation in air.
[16] = 50 M.
Irradiation wavelength was 365 nm.
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(A)
(B)
150
Fluorescence Intensity / a.u.
 10–4
oxalate
/ M–1 ·cm–1
4
3
2
(a)
(c)
1
(b)
250 300
400
500
Wavelength / nm
60
(b)
30
550
600
Wavelength / nm
650
(D)
150
Fluorescence Intensity / a.u.
/ M–1 ·cm–1
(c)
475 500
4
 10–4
90
600
(C)
3
2
(d)
(f)
1
(e)
120
90
(f)
(d)
60
(e)
30
0
0
250 300
9
120
0
0
EDTA
(a)
400
500
Wavelength / nm
600
8
475 500
550
600
Wavelength / nm
650
1
Figure S6. Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 3-carboxymethyllumiflavin
2
16 in an aqueous solution with 35 mM Et3N (closed circles), or 35 mM potassium oxalate (open
3
circles), or 35 mM EDTA (open triangles), or 35 mM DABCO (open squares) in an aqueous solution
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at pH 11 (pH was adjusted with aqueous NaOH).
[T[c,s]T] = 400 M, [16] = 100 M.
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6
50
Recovery / %
40
EDTA
30
NEt3
Oxalate
20
DABCO
10
0
0
7
5
10 15 20 25
Irradiation time / min
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9
10
11
12
13
14
15
9
30
1
Figure S7.
UV spectra of 10ox (a) before and (b) after UV irradiation for 10 min at pH 11 in the
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absence of Et3N at 25°C and UV spectra of 11ox (c) before and (d) after irradiation for 10 min and (e)
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after reoxidation in air.
[10ox] = [11ox] = 50 M.
4
5
/ M–1 ·cm–1
2
 10–4
3
1
+ EDTA
(a)
(b)
0
250 300
6
400
500
Wavelength / nm
7
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9
10
11
12
13
14
10
600
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Figure S8. Change in UV-Vis absorption spectra of 10ox upon addition of 1 mM EDTA in an
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aqueous solution (pH 11) at 25°C: spectra of (a) 10ox and (b) 10ox with 1 mM EDTA.
3
M.
4
5
 10–4
/ M–1 ·cm–1
4
3
(c)
2
(d) and (e)
(a) and (b)
1
0
250 300
6
400
500
Wavelength / nm
7
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9
10
11
12
13
14
11
600
[10ox] = 50
1
Figure S9.
Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 (open squares) and 11
2
(closed squares) in an aqueous solution (pH 11) with 35 mM DABCO in comparison with that by 10 +
3
Et3N (closed circle).
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5
50
Recovery / %
40
10 with Et3N
30
10 with DABCO
11 with DABCO
20
10
0
0
6
2
4
6
8
Irradiation time / min
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10
11
12
13
14
12
10
1
Figure S10. Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 in an aqueous solution
2
with 35 mM EDTA at pH 7.6 (closed circles), or 35 mM EDTA at pH 11 (open circles), or 1 mM
3
EDTA at pH 11 (open triangles), or 35 mM Et3N at pH 11 (open squares).
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5
10ox with 35mM
Et3N (pH 11)
50
10ox with 35 mM
EDTA (pH7.6)
% Recovery
40
30
10ox with 1 mM
EDTA (pH 11)
20
10ox with 35 mM
EDTA (pH 11)
10
0
0
6
2
4
6
8
Irradiation time / min
7
8
9
10
11
12
13
13
10
1
Figure S11. Reaction rates for photoreversion of T[c,s]T (3) at 20°C by 10 in an aqueous solution
2
with 35 mM potassium oxalate at pH 7.6 (closed circles), or 35 mM potassium oxalate at pH 11 (open
3
circles), or 35 mM Et3N at pH 11 (open squares).
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5
50
10ox with 35mM
Et3N (pH 11)
% Recovery
40
30
10ox with 35 mM
oxalate (pH 11)
20
10
0
0
6
2
4
6
8
Irradiation time / min
14
10
10ox with 35 mM
oxalate (pH 7.6)