This journal is © The Royal Society of Chemistry 2000
Supporting Information
for
A Spatially Addressable Combinatorial Library for the Design and Synthesis of Cysteine
Containing Peptides for the Encapsulation of CdS Nanoclusters: Analysis of the Library
Diagonal
Glen Spreitzer, Jacqueline M. Whitling, Jeffry D. Madura and David W. Wright
Department of Chemistry and Biochemistry
Duquesne University
Pittsburgh, PA 15282-1530
S-1. Schematic representation of the spatially addressable combinatorial library containing 125
7-mer peptides XCysYCysZCysGly. X,Y, and Z are the spacer residues -Glu, glutamic acid
with typical linkage, -Glu, -glutamic acid with sidechain linkage, GABA, -aminobutyric acid,
SerGly, serineglycine dipeptide, or -Ahx, -aminohexanoic acid.
S-2. HPLC and mass spectrometry of control peptide.
S-3. HPLC and mass spectrometry of control library peptide.
S-4. Fluorescence spectroscopy of 1-5.
S-5. Representative methyl viologen reduction assay of 2. ---- 2 plus methyl viologen at t = 0
minutes irradiation,
2 plus methyl viologen at t = 10 minutes irradiation, …… 2 plus methyl
viologen at t = 10 minutes irradiation followed by mixing.
S-6. Tabulated data for SEC analysis of peptide/CdS aggregation reaction. Additionally, the
chromatograms of all samples (S-7 to S-11) reveal species attributable to free peptide and low
molecular weight peptide-(Cd2+)n complexes. The molecular weight of the nanoclusters was
estimated from sodium polystyrene sulfonate standards (S-12).
S-7. HPLC trace of ECECECG/CdS reaction mixture. Size exclusion chromatography was
performed with a Waters 600 HPLC system equipped with a Waters 996 photodiode array
detector and a Rheodyne injector using an Ultrahydrogel-250 size-exclusion column (Waters, 7.8
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X 300 mm), variable back pressure (<100 psi), and a flow rate of 0.3 mL/min at 25 oC. The
mobile phase was 0.5 M Tris buffer + 0.075 M KCl. 20 L of reaction mixture was assayed.
S-8. HPLC trace of eCeCeCG/CdS reaction mixture. Conditions as above.
S-9. HPLC trace of gCgCgCG/CdS reaction mixture. Conditions as above.
S-10. HPLC trace of SGCSGCSGCCG/CdS reaction mixture. Conditions as above.
S-11. HPLC trace of hChChCG/CdS reaction mixture. Conditions as above.
S-12. Molecular weight calibration curve using sodium polystyrene sulfonate standards ( 20 L
injection of 15.6-42 M standard, 0.1 M Tris, pH 8.6) having molecular weights of 4,950, 8,000,
16,600, 34,700 and 57,500, respectively. Calibration curve data was obtained under HPLC
conditions as described above.
S-13. CdS sphere with radius of approximately 10 Å shown with a SGCSGCSGCG peptide
ligand bound to the surface.
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S-1.
10
This journal is © The Royal Society of Chemistry 2000
S-2.
HPLC of control peptide, IKDFHVYG-OH (MW 978.1). LCMS identifies the
peak at 4.46 minutes as the peptide.
0
.
8
4
.
4
6
m
i
n
0
.
6
0
.
4
0
.
2
0
.
0
2
4
6
8
1
0
1
2
1
4
T
i
m
e
(
m
i
n
.
)
Ionspray mass spectrometry of control peptide run on a P&E Sciex API III in 0.1%
acetic acid /60% acetonitrile
1
0
0
9
7
7
.
9
8
0
6
0
PercntageofTtalIonCut
4
0
2
0
0
7
0
0
8
0
0
9
0
0
1
0
0
0
1
1
0
0
1
2
0
0
M
o
l
e
c
u
l
a
r
W
e
i
g
h
t
(
D
a
l
t
o
n
)
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This journal is © The Royal Society of Chemistry 2000
S-3.
HPLC of control peptide, hCeCSGCG (mw 770.9). LCMS identifies the
peak at 3.91 minutes as the peptide.
1
.
0
0
.
8
3
.
9
1
m
i
n
.
0
.
6
0
.
4
0
.
2
0
.
0
2
4
6
8
1
0
1
2
1
4
T
i
m
e
(
m
i
n
.
)
Ionspray mass spectrometry of control peptide run on a P&E Sciex API
III in 0.1% acetic acid /60% acetonitrile.
1
0
0
9
0
8
0
7
0
6
0
5
0
7
7
0
.
9
PercntageofTtalIonCut
4
0
3
0
2
0
1
0
0
5
0
0
6
0
0
7
0
0
8
0
0
9
0
0
1
0
0
0
M
o
l
e
c
u
l
a
r
W
e
i
g
h
t
(
D
a
l
t
o
n
)
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This journal is © The Royal Society of Chemistry 2000
S-4.
7
0
0
0
0
6
0
0
0
0
5
0
0
0
0
Counts/Second
4
0
0
0
0
3
0
0
0
0
2
0
0
0
0
1
0
0
0
0
0
3
4
0
3
6
0
3
8
0
4
0
0
4
2
0
4
4
0
4
6
0
4
8
0
5
0
0
W
a
v
e
le
n
g
t
h
(
n
m
)
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This journal is © The Royal Society of Chemistry 2000
S-5.
0
.
8
0
.
7
0
.
6
0
.
5
0
.
4
0
.
3
0
.
2
0
.
1
0
.
0
3
0
0
4
0
0
5
0
0
6
0
0
7
0
0
8
0
0
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S-6.
Peptide
Rt (min)
max (nm)
FWHMb
---306.6 ± 1.8
Size est.
Radius Å
---10.46
---2
M. W.c
(Da)
---30137
ECG*
ECECECG
---23.39 ± 0.034
eCeCeCG
23.61 ± 0.014
320 ± 0.7
10.97
0
28439
gCgCgCG
24.55 ± 0.036
312.1 ± 0.7a
10.66
7.6
22191
SGCSGCSGCG
24.25 ± 0.02
315.3 ± 0.7a
10.79
18.8
24019
hChChCG
25.53 ±0.09
284.1,
9.66,
7.1
17134
24.88 ± 0.07
282.9 ± 1.7 &
319.6 ± 1.2
9.62 & 10.96
4.8 & 5.6
20340
*
Not stable to HPLC SEC analysis.
max shifted when purified by size exclusion chromatography.
b
FWHM = FWHMuv-vis – FWHMhplc
c
Standard deviation of + 5 % from calibration curve.
a
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S-7.
0.12
0.10
0.06
AU
0.08
0.04
0.02
0.00
200.00
300.00
400.00
Wavelength (nm)
0.00
5.00
10.00
15.00
20.00
25.00
30.00
Time (min)
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S-8.
0.14
0.12
0.10
AU
0.08
0.06
0.04
0.02
0.00
200.00
300.00
400.00
0.00
5.00
10.00
15.00
20.00
25.00
Wavelength (nm)
30.00
Time (min)
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This journal is © The Royal Society of Chemistry 2000
S-9.
0.10
0.09
0.08
0.07
0.05
AU
0.06
0.04
0.03
0.02
0.01
0.00
200.00
300.00
400.00
0.00
5.00
10.00
15.00
20.00
25.00
Wavelength (nm)
30.00
Time (min)
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This journal is © The Royal Society of Chemistry 2000
S-10.
0.11
0.10
0.09
0.08
0.06
AU
0.07
0.05
0.04
0.03
0.02
0.01
0.00
200.00
300.00
400.00
0.00
5.00
10.00
15.00
20.00
25.00
Wavelength (nm)
30.00
Time (min)
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S-11.
0.050
0.045
0.040
0.035
0.025
AU
0.030
0.020
0.015
0.010
0.005
0.000
200.00
300.00
400.00
0.00
5.00
10.00
15.00
20.00
25.00
Wavelength (nm)
30.00
Time (min)
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This journal is © The Royal Society of Chemistry 2000
S-12.
4.9
4.7
log MW
4.5
4.3
4.1
3.9
3.7
3.5
20
21
22
23
24
25
26
27
28
29
30
Retention Time (min)
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S-13.
22