SUPPLEMENTARY MATERIAL
Triazole containing zinc(II)dipicolylamine-functionalised peptides as highly
selective pyrophosphate sensors in physiological media
Vincent E. Zwicker,A,† Xuejian Liu,A,† Karen K. Y. Yuen,A Katrina A. JolliffeA,B
A
School of Chemistry, The University of Sydney, 2006, NSW, Australia.
Corresponding author. Email: kate.jolliffe@sydney.edu.au
†These authors contributed equally to this work.
B
Copies of NMR spectra for 1-4.Zn2
Job’s Plots for 1-4.Zn2 with indicator 8
Anion titration data
Composition of Krebs buffer
2
8
8
11
1
2
3
4
5
6
7
Jobs Plots for 1-4.Zn2 with indicator 8
Job’s plot for (left) 1.Zn2 and (right) 2.Zn2 with indicator 8.
Job’s plot for (left) 3.Zn2 and (right) 4.Zn2 with indicator 8.
Results of the titrations of indicator 8 solutions with receptors 1-4.Zn: Batch
2 data at 646
: point 8
100
0.55
80
Intensity
0.45
70
60
0.35
50
40
0.25
30
20
% formation relative to R
Intensity
90
0.4
0.2
10
0.15
0
0
ignored
0.01
0
-0.01
0.004
0.002
0
-0.002
-0.004
250
value
value
Obs-Calc intensity (unweighted)
2
6
10
14
point number
350
450 550
wavelength
650
Left: Absorbance changes for a solution of indicator 8 (20 µM) in Krebs buffer upon addition of 1.Zn2 (0–10 equiv.).
Right: 1:1-fitting curve at 646 nm; inset: residual plot of the residual absorbance at 646 nm vs. the point number.
8
750
: Batch data at 646
: point 8
100
80
Intensity
0.5
70
60
0.4
50
40
0.3
30
20
% formation relative to PV
Intensity
90
0.6
0.6
0.4
0.2
0.2
10
0
0
ignored
0.002
0
-0.002
250
0.005
0
-0.005
value
value
Obs-Calc intensity (unweighted)
2
6
10
14
point number
350
450 550
wavelength
650
750
Left: Absorbance changes for a solution of indicator 8 (20 µM) in Krebs buffer upon addition of 2.Zn2 (0–10 equiv.).
: Batch
at 646
: point 8
Right: 1:1-fitting curve at 646 nm; inset: residual plot of the residual absorbance at
646datanm
vs. the point number.
100
90
80
70
Intensity
0.5
60
50
0.4
40
30
0.3
20
0.2
0.6
% formation relative to R
Intensity
0.6
0.4
0.2
10
0
0
ignored
0.01
0
-0.01
value
value
Obs-Calc intensity (unweighted)
2
6
10
14
point number
0.01
0
-0.01
250
350
450 550
wavelength
650
750
Left: Absorbance changes for a solution of indicator 8 (20 µM) in Krebs buffer upon addition of 3.Zn2 (0–10 equiv.).
Right: 1:1-fitting curve at 646 nm; inset: residual plot of the residual absorbance at 646 nm vs. the point number.
: Batch data at 646
: point 8
100
90
80
Intensity
70
60
50
0.3
40
30
20
% formation relative to R
Intensity
0.6
0.5
0.4
0.2
10
0
Obs-Calc intensity (unweighted)
0.02
0.01
0
-0.01
-0.02
2
6
10
14
point number
0
ignored
0.010
0.005
0
-0.005
-0.010
250
value
value
0.1
350
450 550
wavelength
Left: Absorbance changes for a solution of indicator 8 (20 µM) in Krebs buffer upon addition of 4.Zn2 (0–10 equiv.).
Right: 1:1-fitting curve at 646 nm; inset: residual plot of the residual absorbance at 646 nm vs. the point number.
9
650
750
Results of the titrations of 1:1–receptor:indicator 8 mixtures with PPi, ATP, and ADP for 1.Zn2,
3.Zn2, and 4.Zn2
Left: Absorbance changes of a 1.Zn2:indicator 8 solution (20 µM each) in Krebs buffer upon the addition of PPi (0– 9
equiv.). Right: Changes in the normalised absorbance at 646 nm of the 1:1–1.Zn2:8 mixture upon the addition of PPi,
ATP, and ADP (0–9 equiv.).
Left: Absorbance changes of a 3.Zn2:indicator 8 solution (20 µM each) in Krebs buffer upon the addition of PPi (0– 9
equiv.). Right: Changes in the normalised absorbance at 646 nm of the 1:1–3.Zn2:8 mixture upon the addition of PPi,
ATP, and ADP (0–9 equiv.).
Left: Absorbance changes of a 4.Zn2:indicator 8 solution (20 µM each) in Krebs buffer upon the addition of PPi (0– 9
equiv.). Right: Changes in the normalised absorbance at 646 nm of the 1:1–4.Zn2:8 mixture upon the addition of PPi,
ATP, and ADP (0–9 equiv.).
10
Composition of Krebs buffera
Component
M (g mol-1)
NaCl
58.4
KCl
74.6
CaCl2
111.0
MgSO4.7H2O
246.5
KH2PO4
136.1
NaH2PO4
120.0
Glucose
180.2
TRIS
121.1
a
The pH was adjusted to 7.4 with conc. HCl.
M (g L-1)
8.01
0.40
0.31
0.30
0.054
0.041
1.80
1.21
11
c (mM)
137
5.4
2.8
1.2
0.4
0.3
10
10