Supplementary Material (ESI) for Organic and Biomolecular Chemistry
This journal is © The Royal Society of Chemistry 2007
Supplementary information for
Metal ion induced allosteric transition in the catalytic activity of an artificial
phosphodiesterase
Shinji Takebayashi,a Seiji Shinkai,*a Masato Ikedab and Masayuki Takeuchi*c
a
Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University,
Fukuoka 819-0395, Japan.
b
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering,
Kyoto University, Kyoto 615-8510, JAPAN.
c
Macromolecules Group, Organic Nanomaterials Center, National Institute for Materials Science,
Tsukuba 305-0047, JAPAN.
E-mail: [email protected], [email protected]
Supplementary Material (ESI) for Organic and Biomolecular Chemistry
This journal is © The Royal Society of Chemistry 2007
Zn 0 eq.
N N
a
h
8.40
N
i
8.20
cd
7.75
7.82
g
8.59
h
N
b
N
7.30
g
f
N e d
i
7.58
N N
a
c
b
Zn 2 eq.
N N
N
N
N
N
N N
ppm
9.25
9.00
8.75
8.50
8.25
8.00
7.75
7.50
7.25
7.00
6.75
6.50
Fig. S1. 1H NMR spectra (9.5 ~ 6.5 ppm) for 1+ Zn(ClO4)2; [1]=1.0 mM, pD 7.7 in D2O /
ethanol-d6 = 2 / 1 (v / v) at 5 °C.
Zn 0 eq.
ef
3.81
H
e H
N e
HH
N
N
DPA
L L
Zn 2 eq.
N
N
Zn
e2
e1 f
H e1
e1 H
N
H
H
e2
e2
Jgeminal
Jgeminal
DPA • Zn
ppm
5.75
5.50
5.25
5.00
4.75
4.50
4.25
4.00
3.75
3.50
3.25
3.00
Fig. S2. 1H NMR spectra (6.0 ~ 3.0 ppm) for 1+ Zn(ClO4)2; [1]=1.0 mM, pD 7.7 in D2O /
ethanol-d6 = 2 / 1 (v / v) at 5 °C.
Supplementary Material (ESI) for Organic and Biomolecular Chemistry
This journal is © The Royal Society of Chemistry 2007
(a)
0
600
805.1
823.2
841.2
861.1
879.1
905.2
925.1
943.1
701.2
659.2
1.0
0.5
1007.1
1008.1
N
Zn2+
N N
m / z = 1007.0 (calc.)
741.2
1.5
3(ClO4-)
700
800
900
1007.1
641.3
2.0
1003.1
1004.1
677.1
N
1010.1
1011.2
N
1009.2
1 Zn 2.0 eq.
1005.2
2.5
Intensity / 106cps
+1
N N
Zn2+
N
1006.1
3.0
1100
1000
m/z
1200
1300
(b)
0.6
1271.0
1273.1
1 Zn 8.0 eq.
1274.1
1275.0
1276.0
1277.0
1278.1
1279.0
1270.0
1269.0
N N
Zn2+
1267.0
1268.1
1265.0
1266.0
0.8
5(ClO4-)
0
600
700
800
900
1000
m/z
1100
1230.9
1089.1
1107.1
1143.1
1169.1
1188.8
826.0
0.2
905.2
925.1
943.1
0.4
1206.9
1007.1
m / z = 1270.8 (calc.)
609.0
625.1
Intensity / 106cps
N N
Zn2+
N
N N
Zn2+
N
1271.0
+1
1272.1
1.0
1200
1300
Supplementary Material (ESI) for Organic and Biomolecular Chemistry
This journal is © The Royal Society of Chemistry 2007
(c)
0
600
900
1002.1
1005.1
1003.1
901.1
939.2
865.2
875.0
838.9
803.1
800
1000
m/z
1100
1200
1300
N N
Cu2+
N
N N
Cu2+
N
4.0
1264.0
+1
1 Cu 3.0 eq.
1267.0
1268.1
1268.8
1270.0
1271.1
1266.0
(d)
1261.7
1263.7
5.0
700
821.1
m / z = 1003.0 (calc.)
1.0
0.5
N N
Cu2+
800
1064.1
1084.0
1097.9
700
1003.1
0
600
838.9
865.2
875.0
901.1
1.0
695.5
2.0
900
1000
m/z
1100
1200
1266.0
939.2
m / z = 1265.8 (calc.)
1201.9
1128.0
5(ClO4-)
1138.1
1164.1
3.0
648.1
Intensity / 106cps
N
Cu2+
N N
3(ClO4-)
648.1
676.1
714.1
739.1
757.2
Intensity / 106 cps
N
1006.2
1007.2
N
1.5
1 Cu 2.0 eq.
1004.1
1001.1
1003.1
+1
N N
Cu2+
N
1265.1
2.0
1300
Figure S3. ESI MS spectra for (a) ESI MS spectrum for [Zn2+]/[1] = 2.0 and (b) 8.0 in 33%
ethanol/water (HEPES, 25 mM). ESI MS spectra for (c) [Cu2+]/[1] = 2.0 and (d) 3.0 in 33%
ethanol/water (HEPES, 25 mM).
Supplementary Material (ESI) for Organic and Biomolecular Chemistry
This journal is © The Royal Society of Chemistry 2007
2.5
(a)
4.0
(b)
N N
N N
N
N
N
N
(dc/dt)-1 / 107 M-1s
(dc/dt)-1 / 107 M-1s
2.0
N
N
N N
1.5
1.0
N N
N N
0.5
N
N N
0.5
1.0
1.5
[HPNP]-1 / 103 M-1
N N
N N
N
N
2.0
N N
N N
N
N
and
N
1.0
N
N
N
N
N
N
N
N N
N N
N
0
0
N
N
N
N
0
3.0
2.0
2.5
0
1.0
2.0
3.0
[HPNP]-1 / 103 M-1
4.0
5.0
Figure S4. Lineweaver-Burk plots for HPNP cleavage; in 33% ethanol/water (HEPES, 25 mM),
pH 7.7 at 25 °C; (a) [1] = 0.4 mM, [Zn(ClO4)2] = 0.80 mM (solid line), [1] = 0.40 mM, [Zn(ClO4)2]
= 2.0 mM (broken line), (b) [1] = 1.0 mM, [Cu(ClO4)2] = 2.0 mM (solid line), [1] = 1.0 mM,
[Cu(ClO4)2] = 3.0 mM (broken line).
(a)
EDTA-2Na 0 ~ 3.0 eq.
(b) EDTA 3.0 ~ 6.0 eq.
1.2
1.2
0 eq.
1.0
0.8
3.0 eq.
0.6
0.4
0.8
3.0 eq.
0.6
0.4
6.0 eq.
0.2
0.2
0
Absorbance
Absorbance
1.0
260
280 300 320
Wave length / nm
340
0
260
280 300 320
Wave length / nm
340
Fig. S5 UV-Vis spectral changes of 1•(Zn2+)3 ([1] = 0.40 mM, [Zn(ClO4)2]) upon addition of
EDTA-2Na ((a) 0-3 equiv., (b) 3-6 equiv.) in 33% ethanol/water (HEPES, 25 mM) at pH 7.7 and
25 °C.