1

Electronic Supplementary Information

A novel fluorescent peptidyl probe for highly sensitive and selective ratiometric detection of Cd(II) in aqueous and bio-samples via metal ion-mediated self-assembly

Kwan Ho Jung,a Semin Oh,a Joohee Park,a Yu Jin Park,a See-Hyoung Park,*,b and Keun-Hyeung Lee*,a

aCenter for Design and Applications of Molecular Catalysts, Department of Chemistry and Chemical Engineering, Inha University, Incheon, 402-751, South Korea. bDepartment of Bio and Chemical

Engineering, Hongik University, Sejong 30016, Republic of Korea E-mail: leekh@inha.ac.kr (K. –H. Lee)

Contents

Scheme S1. Synthesis scheme of 1 S2

Figure S1. HPLC chromatogram of 1 S3

Figure S2. ESI-Mass spectrum of 1 S4

Figure S3. 1H NMR of 1

Figure S4. 13C NMR of 1

Figure S5. Solvent effect on fluorescence emission spectra of 1 with Cd2+

Figure S6. Intensity ratio change of 1 with Cd2+

Figure S7. Emission intensity ratio of 1 induced by various sources of Cd2+

Figure S8. Interference effect of metal ion on the ratiometric response to Cd2+

Figure S9. Florescent spectra of 1 with Cd2+ in aqueous solutions containing 3% DMF

Figure S10. Job’s Plot for 1 with Cd2+

Figure S11. Non-linear least square fitting of the intenisty as a fucntion of cocentration of Cd2+

Figure S12. Reversibility of 1 for Cd2+

Figure S13. CD Spectra of 1 in the presence and absence of Cd2+

Figure S14. Linear curve fitting of intensity ratio as a function of Cd2+ in urine samples

Figure S15. Ratiometric response to Cd2+ in ground water samples

Figure S16. Stability performance of 1

Figure S17. Toxicity study of 1 for cells

Table S1. Comparison of the properties of ratiometric fluorescent probes for Cd(II)

References

S5

S6

S7

S8

S9

S10

S11

S12

S13

S14

S15

S16

S17

S18

S19

S20

S21

Electronic Supplementary Material (ESI) for New Journal of Chemistry.This journal is © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2018

2

N

NH

HO

O

O

HS

O

NH

OHN

SHO

Cli) Swelling in DCMii) Fmoc-Cys(Trt)-OH (5 equiv)

DIPEA (6 equiv) in DCM : DMF (1:1))ii) DCM : MeOH : DIPEA (17:2:1) 20 mini) Swlling in DMF

iii) DCM : MeOH : DIPEA (17:2:1) 5 min

i) 25% piperidine in DMFii) Fmoc-Pro-OH (3 equiv)

HOBt (3 equiv), DIC (3 equiv)

i) 25% piperidine in DMFii) Fmoc-Gly-OH (3 equiv)

HOBt (3 equiv), DIC (3 equiv)

i) 25% piperidine in DMFii) Fmoc-Cys(Trt)-OH (3 equiv)

HOBt (3 equiv), DIC (3 equiv)

TFA : TIS : Water(95 : 2.5 : 2.5)

i) 25% piperidine in DMFii) Pyrene acetic acid (3 equiv)

HOBt (3 equiv), DIC (3 equiv)

Cl

Scheme S1. Synthetic scheme of 1.

3

Figure S1. HPLC chromatogram of 1.

4

[M + H+]+

Calculated mass: 621.18

Figure S2. ESI-Mass spectrum of 1.

5

Figure S3. 1H NMR of 1.

6

Figure S4. 13C NMR of 1.

7

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

1% DMF 10% DMF 25% DMF

50% DMF 75% DMF 100% DMF

400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 1 + Cd(II)

Figure S5. Fluorescence emission spectra of 1 (15 μM) with Cd2+ (15 μM) in aqueous buffered solutions (10 mM, HEPES, pH 7.4) containing various volume of DMF.

8

0 5 10 15 200

10

20

30

I 475/I 39

5

[Cd(II)] (M)0 5 10 15 20

0

1

2

3

I 475/I 43

3

[Cd(II)] (M)

(a) (b)

0 5 10 15 200

1

2

3

4

I 475/I 43

5

[Cd(II)] (M)

(b)

Figure S6. Intensity ratio change (a, I475/I395; b, I475/I433) of 1 (15 μM) as a function of Cd2+ in aqueous buffered solution (10 mM HEPES, pH 7.4) containing 1% DMF (λex = 342 nm).

9

None Cd(ClO4)2 CdCl2 Cd(NO3)20

4

8

12

16

20

24

28

32

I 475/I 39

5

Figure S7. Emission intensity ratio of 1 (10 μM) induced by various sources of Cd2+ (1 equiv) in aqueous buffered solution (10mM HEPES, pH 7.4) containing 1% DMF .

10

None Ag(I) Al(III) Ca(II) Co(II) Cr(III) Cu(II) Fe(II) Fe(III) Hg(II) K(I) Mg(II) Mn(II) Na(I) Ni(II) Pb(II) Zn(II) --0

5

10

15

20

25

30

35

I 475 /

I 395

Cd(II) + each metal ions

Figure S8. Emission intensity ratio of 1 (15 μM) in the presence of Cd2+ (1 equiv) and various metal ions (5 equiv) in aqueous buffered solution (10mM HEPES, pH 7.4) containing 1% DMF.

11

350 400 450 500 550 6000

200

400

600

800

Inte

nsity

Wavelength (nm)

1 (75M) 1 + Cd2+ (1 equiv)

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength (nm)

1 (150M) 1 + Cd2+ (1 equiv)

(a)

(b)

Figure S9. Fluorescence emission spectra of 1 (a; 75 μM, b; 150 μM) in the absence and presence of Cd2+ (1 equiv) in aqueous buffered solutions (2 mM, HEPES, pH 7.4) containing 3% DMF (λex = 342 nm).

12

0.0 0.2 0.4 0.6 0.8 1.0

0

50

100

150

200

250

300

I 470

[Cd(II)] / [Cd(II)] + [Probe]

Figure S10. Job’s plot for 1 with Cd2+ in aqueous buffered solution (10 mM HEPES, pH 7.4) containing 1% DMF; total concentration = 15 μM, slit 15/12 nm, 1% attenuator (λex = 342 nm).

13

0 5 10 15 200

200

400

600

I 475

[Cd(II)] (M)

Equation y = P4+P3*((P1+x+P2)-sqrt(((P1+x+P2)^2)-4*P1*x))/2

Adj. R-Square 0.99679Value Standard Error

B P1 15 0B P2 1.3042E-4 0.00198B P3 35.40695 0.25872B P4 1.1 0

Figure S11. Non-linear least square fitting of the emission intenisty of 1 (15 μM) as a fucntion of cocentration of Cd2+ by a 1:1 complex model.

14

350 400 450 500 550 6000

200

400

600

800

1000

Inte

nsity

Wavelength(nm)

1 1 + Cd2+

1 + Cd2+ + EDTA

Figure S12. Fluorescence emission spectra of 1 (10 μM) in the absence and presence of Cd2+ (1 equiv) and EDTA (1 equiv) in aqueous buffered solutions (10 mM, HEPES, pH 7.4) containing 1% DMF (λex = 342 nm).

15

280 300 320 340 360 380 400-2.5-2.0-1.5-1.0-0.50.00.51.01.52.02.5

285 nmCD

(mde

g)

Wavelength (nm)

11 + Cd(II)

355 nm

190 200 210 220 230 240 250 260 270-20-16-12-8-4048

121620

CD

(mde

g)

Wavelength (nm)

11 + Cd(II)

(a) (b)

Figure S13. (a) Far- and (b) Near-UV CD Spectra of 1 (75 μM) in the absence or presence of Cd2+ (75 μM) in aqueous buffered solutions (10 mM PBS, pH 7.4) containing 5% (v/v) 2,2,2-trifluoroethanol.

16

0 50 100 150 200 250 300 3500.240.270.300.330.360.390.420.450.48 Fi = 0.0005 conc(nM) + 0.279

R2 = 0.974

I 475 /

I 447

[Cd(II)] (nM)0 50 100 150 200 250

0.10

0.11

0.12

0.13

0.14

0.15

0.16

I 475 /

I 433

Fi = 0.0002 conc(nM) + 0.104R2 = 0.984

[Cd(II)] (nM)

(a) (b)

Figure S14. Linear curve fitting of emission intensity ratio change of 1 (15 μM) as a function of the concentration of Cd2+ in aqueous buffered solutions (10 mM, HEPES, pH 7.4) containing urine samples.

17

350 400 450 500 550 600

0

200

400

600

800

1000

In

tens

ity

Wavelength (nm)

0 5 10 15 20 25

0

200

400

600

800

1000 I395

I475

Inte

nsity

Concentration (M)

Figure S15. Emission spectra of 1 (15 μM) with increasing concentration of Cd2+ in aqueous buffered solutions (10 mM HEPES, pH 7.4) containing ground waters.

18

0 5 10 15 20 25

0

5

10

15

20

25

30

1 1 + Cd(II)

I 475/I 39

5

Time (hour)

(a)

0 20 40 60 80 100

0

5

10

15

20

25

30

1 1 + Cd(II)

I 475/I 39

5

Time (min)

(b)

Figure S16. (a) Incubation of stock solution of 1 at room temperature for 24 hrs and emission intensity ratio of 1 (15 µM) by Cd2+ (15 µM) (b) Upon addition of Cd2+ into the solution containing 1, emission intesnity ratio induced by Cd2+ for 100 mins in aqueous buffered solutions (10 mM, HEPES, pH 7.4) containing 1% DMF.

19

Cd(II): 15 uM

Py: 30 uM

EDTA: 150 uM

- - -

+ - -

+ - +

- + -

+ + -

+ + +

0 20 40 60 80 100 120Cell Viability (%)1

Figure S17. MTS assay for the viability of MDA-MB-231 cells in DMEM 10% FBS treated with 1, 1 + Cd(ClO4)2, and 1 + Cd(ClO4)2 + EDTA for 24 h.

20

Table S1 Comparison of the properties of ratiometric fluorescent probes for Cd(II) in aqueous solution.1-11

aND means not determined.

Fluorophore(s) Organic cosolvent

Emission bands (nm)

Change (fold) LOD Response Application

DansylTrp 0% 350 to

500 9 0.9M Hg(II), Zn(II), Ag(I) No cell image

DansylTrp 0% 350 to

500 4 0.3M Cu(II), Zn(II) No cell image

5-Dimethylamino-2-(2-pyridinyl)-

benzoimidazole)0% 493 to

587 8 0.3 pM Zn(II) Cell image

Coumarin 0% 328 to 368 3.5 40 pM Zn(II) Cell image

4,5-Diamino-1,8-naphthalimide 10% EtOH 487 to

531 3 0.1 M Zn(II) No cell image

Boradiazaindacene(BODIPY)

90% Acetone

550 to 800 13 NDa Cr(III), Ni(II), Cu(II) Cell image

8-Hydroxyquinolinenorbornene

50% Methanol

330 to 600 2 1.6 nM Zn(II) Paper strip

8-Hydroxyquinoline 80% Ethanol

350 to 650 4.5 23.6 nM Zn(II) Cell image

8-Hydroxy-2-methylquinoline

80% Dioxane

400 to 700 92 20 nM Cu(II), Zn(II) No cell

image4-Isobutoxy-6-

(dimethylamino)-8-methoxyquinaldine

0% 400 to 700 3 9.6 pM

Mn(II),Fe(II),Co(II),Ni(II), Cu(II), Hg(II), Pb(II),

Zn(II)Cell image

Phenanthro[9,10-d]oxazole 50% DMF 400 to

600 NDa NDa Fe(III), Hg(II),Pb(II),Cu(II) Paper strip

Pyrene(Present work) 1% DMF 395 to

475 28 22 nM Only Cd(II) Cell image, Urine

References 1 B. P. Joshi, J. Park, W. I. Lee, K. H. Lee, Talanta., 2009, 78, 903−909.2 Y. Li, L. Li, X. Pu, G. Ma, E. Wang, J. Kong, Z. Liu, Y. Liu, Bioorg. Med. Chem. Lett., 2012, 22,

4014−4017.3 Z. Liu, C. Zhang, W. He, Z. Yang, X. Gao, Z. Guo, Chem. Commun., 2010, 46, 6138−6140.4 M. Taki, M. Desaki, A. Ojida, S. Iyoshi, T. Hirayama, I. Hamachi, Y. Yamamoto, J. Am. Chem. Soc.,

2008, 130, 12564−12565.5 C. Lu, Z. Xu, J. Cui, R. Zhang, X. Qian, J. Org. Chem., 2007, 72, 3554−3557.6 X. Peng, J. Du, J. Fan, J. Wang, Y. Wu, J. Zhao, S. Sun, T. Xu, J. Am. Chem. Soc., 2007, 129,

1500−1501.7 S. Sarkar, R. Shunmugam, ACS Appl. Mater. Interfaces., 2013, 5, 7379−7383.8 Z. Shi, Q. Han, L. Yang, H. Yang, X. Tang, W. Dou, Z. Li, Y. Zhang, Y. Shao, L. Guan, W. Liu,

Chem. Eur. J., 2015, 21, 290−297.9 Y. Bao, B. L. Liu, H. Wang, F. Du, R. Bai, Anal. Methods., 2011, 3, 1274−1276.10 L. Xue, G. Li, Q. Liu, H. Wang, C. Liu, X. Ding, S. He, H. Jiang, Inorg. Chem., 2011, 50, 3680−3690.11 L. Ahang, Q. Tong, L. Shi, Dalton Trans., 2013, 42, 8567−8570.