![Page 1: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/1.jpg)
SUPPLEMENTARY INFORMATIONModulation of i-Motif Thermal Stability by Insertion of Anthraquinone Monomers
Alaa S. Gouda a, b, Mahasen S. Amine b and Erik B. Pedersen*a
aDepartment of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, 5230
Odense M, Denmark. Fax +45 66158780; *E-mail: [email protected] Department of Chemistry, Faculty of Science, Benha University, Benha, Egypt13518.
Circular Dichroism (CD) spectral analysis of human telomeric DNA (ON1) and
anthraquinone-modified i-motif variants (ON2-ON33, Fig. S1-S3):
Figure S1. Circular dichroism (CD) spectra of human telomeric i-motif DNA (ON1) and anthraquinone-modified i-motif variants (ON2-ON13) measured at 20 ºC. For sample composition see table 1.
380.2360.2340.2320.2300.2280.2260.2240.2220.2200.2-6
-4
-2
0
2
4
6
8
10
12ON1ON2ON3ON4ON5ON6ON7ON8ON9ON10ON11ON12ON13
Wavelength / nm
CD
/ m
deg
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2017
![Page 2: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/2.jpg)
380.2360.2340.2320.2300.2280.2260.2240.2220.2200.2-6
-4
-2
0
2
4
6
8
10ON1ON30ON31ON32ON33
Wavelength / nm
CD
\ mde
g
380.2360.2340.2320.2300.2280.2260.2240.2220.2200.2-6
-4
-2
0
2
4
6
8
10ON1ON14ON15ON16ON17ON18ON19ON20ON21ON22ON23ON24ON25ON26ON27ON28ON29
Wavelength / nm
CD
\ m
deg
Figure S2. CD spectra of human telomeric i-motif DNA (ON1) and anthraquinone-modified i-motif variants (ON14-ON29) measured at 20 ºC. For sample composition see table 1.
Figure S3. CD spectra of human telomeric i-motif DNA (ON1) and anthraquinone-modified i-motif variants (ON30-ON33) measured at 20 ºC. For sample composition see table 1.
![Page 3: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/3.jpg)
25 30 35 40 45 50 55 60 65-0.1
-0.08
-0.06
-0.04
-0.02
0
0.02
0.04ON1ON2ON3ON4ON5ON6ON7ON8ON9ON10ON11ON12ON13
Temperature (ºC)
dA29
5nm
/dT 25 30 35 40 45 50 55 60 65
-0.1
0.1
0.3
0.5
0.7
0.9
1.1
ON1ON2ON3ON4ON5ON6ON7ON8ON9ON10ON11ON12ON13
Temperature (ºC)
Abs
orba
nce
Normalized UV absorption melting curves and first derivatives of human telomeric i-motif
DNA (ON1) and anthraquinone-modified i-motif variants (ON2-ON33, Fig S4-S6):
Figure S4. Normalized UV absorption melting curves (Up) and first derivatives (Down) versus temperature difference (0.5 ºC/min) from 25-65 ºC for i–motifs (ON1-ON13) at 295 nm (4 µM) in potassium buffer (100 mM KCl + 20 mM K2HPO4 (pH= 5.5) and 1 mM K2EDTA). Melting temperatures are within the uncertainty ± 0.5 °C as determined by repetitive experiments and Tm values were calculated taking an average of the two melting curves.
![Page 4: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/4.jpg)
25 30 35 40 45 50 55 60 65-0.09
-0.07
-0.05
-0.03
-0.01
0.01
ON1ON14ON15ON16ON17ON18ON19ON20ON21ON22ON23ON24ON25ON26ON27ON28ON29
Temperature (ºC)
dA 2
95nm
/dT
25 30 35 40 45 50 55 60 65-0.1
0.1
0.3
0.5
0.7
0.9
1.1ON1ON14ON15ON16ON17ON18ON19ON20ON21ON22ON23ON24ON25ON26ON27ON28ON29
Temperature (ºC)
Abs
orba
nce
25 30 35 40 45 50 55 60 65-0.1
0.1
0.3
0.5
0.7
0.9
1.1ON1ON30ON31ON32ON33
Temperature (ºC)
Abs
orba
nce
Figure S5. Normalized UV absorption melting curves (Up) and first derivatives (Down) versus temperature difference (0.5 ºC/min) from 25-65 ºC for i–motifs (ON1 and ON14-ON29) at 295 nm (4 µM) in potassium buffer (100 mM KCl + 20 mM K2HPO4 (pH= 5.5) and 1 mM K2EDTA). Melting temperatures are within the uncertainty ± 0.5 °C as determined by repetitive experiments and Tm values were calculated taking an average of the two melting curves.
![Page 5: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/5.jpg)
25 30 35 40 45 50 55 60 65-0.09
-0.08
-0.07
-0.06
-0.05
-0.04
-0.03
-0.02
-0.01
0
0.01
ON1ON30ON31ON32ON33
Temperature (ºC)
dA 2
95nm
/dT
Figure S6. Normalized UV absorption melting curves (Up) and first derivatives (Down) versus temperature difference (0.5 ºC/min) from 25-65 ºC for i–motifs (ON1 and ON30-ON33) at 295 nm (4 µM) in potassium buffer (100 mM KCl + 20 mM K2HPO4 (pH= 5.5) and 1 mM K2EDTA). Melting temperatures are within the uncertainty ± 0.5 °C as determined by repetitive experiments and Tm values were calculated taking an average of the two melting curves.
Table S1. MALDI-TOF mass spectrometry data (calculated and found mass) of i-Motifs
![Page 6: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/6.jpg)
Oligo bases replaced Sequence Mcalcd Mfound
ON1 wild type 5'-CCC TAA CCC TAA CCC TAA CCC T-3' 6504.44 6504.13ON2 T10 5'-CCC TAA CCC H18AA CCC TAA CCC T-3' 6617.43 6618.68ON3 A11 5'-CCC TAA CCC TH18A CCC TAA CCC T-3' 6608.43 6608.07ON4 A12 5'-CCC TAA CCC TAH18 CCC TAA CCC T-3' 6608.43 6609.25ON5 T10 5'-CCC TAA CCC H14AA CCC TAA CCC T-3' 6617.43 6617.38ON6 A11 5'-CCC TAA CCC TH14A CCC TAA CCC T-3' 6608.43 6608.94ON7 A12 5'-CCC TAA CCC TAH14 CCC TAA CCC-T-3' 6608.43 6608.07ON8 T10 5'-CCC TAA CCC H15AA CCC TAA CCC T-3' 6617.43 6618.29ON9 A11 5'-CCC TAA CCC TH15A CCC TAA CCC T-3' 6608.43 6609.22
ON10 A12 5'-CCC TAA CCC TAH15 CCC TAA CCC T-3' 6608.43 6609.30ON11 T10 5'-CCC TAA CCC H26AA CCC TAA CCC T-3' 6617.43 6617.49ON12 A11 5'-CCC TAA CCC TH26A CCC TAA CCC T-3' 6608.43 6608.65ON13 A12 5'-CCC TAA CCC TAH26 CCC TAA CCC-T-3' 6608.43 6608.36ON14 A6T16 5'-CCC TAH18 CCC TAA CCC H18AA CCC T-3' 6721.56 6722.74ON15 A6T16 5'-CCC TAH14 CCC TAA CCC H14AA CCC T-3' 6721.56 6722.85ON16 A6T16 5'-CCC TAH15 CCC TAA CCC H15AA CCC T-3' 6721.56 6728.08ON17 A6T16 5'-CCC TAH26 CCC TAA CCC H26AA CCC T-3' 6721.56 6721.66ON18 A6T16 5'-CCC TAH18 CCC TAA CCC H14AA CCC T-3' 6721.56 6723.44ON19 A6T16 5'-CCC TAH18 CCC TAA CCC H15AA CCC T-3' 6721.56 6725.84ON20 A6T16 5'-CCC TAH18 CCC TAA CCC H26AA CCC T-3' 6721.56 6721.66ON21 A6T16 5'-CCC TAH14 CCC TAA CCC H18AA CCC T-3' 6721.56 6722.85ON22 A6T16 5'-CCC TAH14 CCC TAA CCC H15AA CCC T-3' 6721.56 6724.04ON23 A6T16 5'-CCC TAH14 CCC TAA CCC H26AA CCC T-3' 6721.56 6722.76ON24 A6T16 5'-CCC TAH15 CCC TAA CCC H18AA CCC T-3' 6721.56 6723.74ON25 A6T16 5'-CCC TAH15 CCC TAA CCC H14AA CCC-T-3' 6721.56 6722.26ON26 A6T16 5'-CCC TAH15 CCC TAA CCC H26AA CCC T-3' 6721.56 6722.18ON27 A6T16 5'-CCC TAH26 CCC TAA CCC H18AA CCC T-3' 6721.56 6719.88ON28 A6T16 5'-CCC TAH26 CCC TAA CCC H14AA CCC T-3' 6721.56 6722.26ON29 A6T16 5'-CCC TAH26 CCC TAA CCC H15AA CCC T-3' 6721.56 6718.12ON30 A18 5'-CCC TAA CCC TAA CCC TAH18 CCC T-3' 6608.43 6608.28ON31 A18 5'-CCC TAA CCC TAA CCC TAH14 CCC-T-3' 6608.43 6612.98ON32 A18 5'-CCC TAA CCC TAA CCC TAH15 CCC T-3' 6608.43 6608.36ON33 A18 5'-CCC TAA CCC TAA CCC TAH26 CCC T-3' 6608.43 6609.83
Maldi-TOF Mass Spectrometry Data of Oligonucleotides (ON1-ON33)
![Page 7: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/7.jpg)
ON1
ON2
ON3
![Page 8: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/8.jpg)
![Page 9: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/9.jpg)
ON4
ON5
![Page 10: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/10.jpg)
ON6
ON7
![Page 11: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/11.jpg)
ON8
ON9
![Page 12: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/12.jpg)
ON10
ON11
ON1
![Page 13: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/13.jpg)
ON12
ON13
![Page 14: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/14.jpg)
ON14
ON15
![Page 15: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/15.jpg)
ON16
ON17
![Page 16: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/16.jpg)
ON18
ON19
![Page 17: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/17.jpg)
ON20
ON21
![Page 18: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/18.jpg)
ON22
ON23
![Page 19: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/19.jpg)
ON24
ON25
![Page 20: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/20.jpg)
ON26
ON27
![Page 21: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/21.jpg)
ON28
ON29
![Page 22: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/22.jpg)
ON30
ON31
![Page 23: SUPPLEMENTARY INFORMATION · 2017. 7. 20. · ON10 ON11 ON12 ON13 Temperature (ºC) dA 295nm /dT 25 30 35 40 45 50 55 60 65-0.1 0.1 0.3 0.5 0.7 0.9 1.1 ON1 ON2 ON3 ON4 ON5 ON6 ON7](https://reader033.vdocuments.site/reader033/viewer/2022061003/60b20fd277a035659211f378/html5/thumbnails/23.jpg)
ON32
ON33