Rationally Designed Fluorescence Turn-On Sensor for Cu 2+

Transcription

1 Supporting Information for Rationally Designed Fluorescence Turn-On Sensor for Cu 2+ Kyoung Chul Ko, a Jia-Sheng Wu, b Hyun Jung Kim, b Pil Seung Kwon, c Jong Wan Kim, c Richard A. Bartsch, d Jin Yong Lee, a, * and Jong Seung Kim b, * a Department of Chemistry, Sungkyunkwan University, Suwon Republic of Korea. jinylee@skku.edu b Department of Chemistry, Korea University, Seoul , Republic of Korea. Fax: (+) ; Tel: ; jongskim@korea.ac.jr c Department of Laboratory Medicine, Dankook University Hospital, Cheonan , Republic of Korea. d Department of Chemistry and Biochemistry,Texas Tech University, Lubbock, Texas 79401, USA S1

2 Table of Contents Page Calculation detail S3 Figure S S4 Figure S S4 Figure S S5 Figure S S5 Figure S S6 Figure S S6 Figure S S9 Figure S S9 Figure S S10 Figure S S10 Figure S S11 Figure S S11 Figure S S12 Figure S S12 Figure S S13 Figure S S13 Table S S14 Cell Incubation Procedure S14 Scheme S S15 Synthesis of IC S15 References of SI S16 S2

3 Calculation detail All geometry optimizations were performed using Becke s three-parameter B3LYP exchange-correlation functional and the 6-31G* basis set. We firstly compared the stabilities of the two structures (crystal structure and proposed structure in Figure S5) of IC in both the gas and solution phases. For the solution phase, we employed PCM calculations using water as solvent because a mixed solvent is currently not available, though a 1:4 mixture of CH 3 C and water was used in the experiment. To determine the optimized structure of IC-Cu 2+, we should verify determined the most probable geometry of IC-Cu 2+ by comparing the diverse optimized structures. Because a water molecule also can bind with Cu 2+ as a ligand, we add a water molecule to IC-Cu 2+ complex one by one. As a result, IC and Cu 2+ with two water molecules might form a most probable structure of IC-Cu 2+ complex. When three water molecules were used as ligands, the complex has almost same geometry that of complex with two water molecules. And the latest added water molecule migrates to makes a hydrogen bonding between the water molecules and does not make the interaction with Cu 2+ at all (Figure S6). In addition, our recent report also can support that this complex geometry can represent the appropriate conformation of IC-Cu 2+. S1 At the optimized geometry, we obtained the excitation properties such as excitation energies and the contribution of relevant orbital transitions to the excitation by the TDDFT calculations. S3

4 0.6 Absorbance [Cu 2+ ] 0 eq 0.3 eq 0.5 eq 0.7 eq 1 eq 10 eq 50 eq Wavelength (nm) Figure S1. UV-Vis spectra of IC (10 µm) in 80% aqueous MeC 1:4 (PBS buffer solution, ph=7.2) with different concentrations of CuCl 2. Figure S2. Job s plot of a 1:1 complex of IC and Cu 2+, where the difference in fluorescence intensity at 520 nm is plotted against the mole fraction of IC at an invariant total concentration of 3.0 μm in 80% aqueous MeC (PBS buffer solution, ph = 7.2). S4

5 Voyager Spec #1[BP = 274.2, 26937] % Intensity Mass (m/z) Figure S3. MALDI-TOF MS spectrum of IC and Cu 2+ adduct in MeC. Figure S4. Fluorescence intensity at 515 nm (excitation at 460 nm) of IC (3.0 μm) and IC + 25 equiv of Cu 2+ in 80% aqueous MeC with different ph conditions. S5

6 Figure S5. Optimized structures of IC; CS (a) and BS (b), and calculated stability of IC in the gas phase and water solvent. Figure S6. (a) Optimized structure of IC-Cu 2+ (b) Optimized structure of IC-Cu 2+ with one H 2 O (as a ligand) (c) Optimized structure of IC-Cu 2+ with two H 2 Os (d) Optimized structure of IC-Cu 2+ with three H 2 Os. S6

7 IC Molecular Orbital Orbital of selected nitrogen atom LUMO HOMO HOMO-1 HOMO-2 IC-Cu 2+ -2H 2 O Molecular Orbital Orbital of selected nitrogen atom LUMO (alpha) HOMO (alpha) HOMO-1 (alpha) S7

8 HOMO-2 (alpha) HOMO-3 (alpha) HOMO-7 (alpha) LUMO+1 (beta) LUMO (beta) HOMO (beta) HOMO-1 (beta) S8

9 HOMO-3 (beta) HOMO-6 (beta) HOMO-8 (beta) Figure S7. Calculated molecular orbitals of IC and IC-Cu 2+. S2 Figure S8. Molecular orbitals of nitrogen atom for IC. S2 S9

10 Figure S9. Molecular orbitals of nitrogen atom for IC-Cu 2+. Fluorophore orbital itrogen lone-pair orbital hν hν hν' Cu 2+ IC IC-Cu 2+ Figure S10. Proposed mechanism of fluorescence enhancement. S10

11 Figure S11. Fluorescence spectra of the control mouse urine collected over 24hs (green), the Cu 2+ -fed mouse urine with IC added (orange), and the Cu 2+ -fed mouse urine (pink) after addition of IC. Figure S12. 1 H MR (DMSO-d 6, 200 MHz) spectrum of IC. S11

12 Figure S C MR (DMSO-d 6, 50 MHz) spectrum of IC. Figure S14. FAB MS spectrum of IC in MeC. S12

13 Figure S15. 1 H MR data of IC (10 mm) in DMSO-d 6 in the presence and absence of 0.3 equiv of Cu(ClO 4 ) Fluorescence Intensity (a.u.) R= Concentration (10-6 M) Figure S16. The correlation between the fluorescence intensity and Cu 2+ concentration. S13

14 Table S1. Calculated electronic contributions of excitations for IC and IC-Cu 2+. Theoretical λ max (nm) IC 386 IC-Cu Contributions of Excitation HOMO-->LUMO (3.4%) HOMO-1-->LUMO (94.0%) HOMO-2-->LUMO (2.6%) HOMO-2 (α)-->lumo(α) (1.8%) HOMO-1(α)-->LUMO(α) (44.9%) HOMO(α)-->LUMO(α) (1.7%) HOMO-6(β)-->LUMO(β) (1.9%) HOMO-3(β)-->LUMO+1(β) (1.8%) HOMO-1(β)-->LUMO+1(β) (42.6%) HOMO(β)-->LUMO+1(β) (5.2%) Cell Incubation Procedure. A LLC-MK2 cell line was prepared for fluorescence imagining. Cell lines from continuous cultures in Dulbeccos-modified Eagles medium (GibcoBRL, USA), supplemented with 10% (v/v) heat-inactivated fetal calf serum (HyClone), ug/ml of penicillin, ug/ml of streptomycin and 0.25 mm ICglutamine, at 37 o C in 5% CO 2 humidified air. When the cells reached the logarithmic phase, the cell density was adjusted to cells per well in cell media. The cells were then used to inoculate in a glass bottom dish (Precision Instruments Inc, USA) with 1.0 ml of cell suspension in each well. After cell adhesion, the culture medium was removed. The cell layer was rinsed twice with phosphate buffered saline (PBS), and then 1.0 ml of culture medium was placed in each well. S14

15 CHO 1) CH 2 (COOC 2 H 5 ) 2 POCl 3 OH 2) HCl/AcOH O O DMF 1 O CHO O H 2 SH O O SH 2 IC Scheme S1. Synthetic pathway of IC. Synthesis of IC 7-Diethylamino coumarin-3-aldehyde (2) was prepared according to previous report. S3 A portion of 7-diethylamino coumarin-3-aldehyde (245.0 mg, 1.0 mmol) and 4-amino-5- phenyl-4h-1,2,4-triazole-3-thiol (222.4 mg, 1.10 mmol) were combined in hot absolute ethanol (20.0 ml) for 2h to yield a scarlet precipitate. The solution was stirred at reflux conditions for 4 h, and the precipitate was filtrated, washed with hot absolute ethanol 3 times, then recrystallized with CHCl 3 /C 2 H 5 OH (v/v, 1/3) to obtain the yellow crystals of IC (366.4 mg, 0.85 mmol) in 85 % yield. Mp 186 ~ 187 C. IR (KBr pellet, cm -1 ): 1698, 1616, 603; 1 H-MR (DMSO-d 6, 200 MHz) δ (s, 1H), 9.56 (s, 1H), 8.54 (s, 1H), (m, 2H), (d, J = 9.3Hz, 1H), (s, 3H), (d, J = 9.1Hz 1H), 6.59 (s, 1H), 3.34 (s, 4H), 1.15 (t, J = 6.79Hz, 6H). 13 C-MR (DMSO-d 6, 50 MHz) δ 162.3, 160.0, 157.8, 152.7, 148.5, 142.9, 138.7, 132.0, 130.6, 128.7, 128.3, 125.6, 110.2, 109.4, 180.0, 96.4, 44.4, 40.7, 40.3, 39.9, 39.5, 39.1, 38.6, 38.2, 12.4 ppm. FAB MS m/z (M + ): calcd, Found, S15

16 References of SI S1. H. S. Jung, P. S. Kwon, J. W. Lee, J. I. Kim, C. S. Hong, J. W. Kim, S. H. Yan, J. Y. Lee, J. H. Lee, T. Joo and J. S. Kim, J. Am. Chem. Soc., 2009, 131, S2. The orbital figures were drawn with POSMOL package. S. J. Lee, H. Y. Chung, K. S. Kim, Bull. Korean Chem. Soc., 2004, 25, S3. J.-S. Wu, W.-M. Liu, X.-Q. Zhuang, F. Wang, P.-F. Wang, S.-L. Tao, X.-H. Zhang, S.- K. Wu, S.-T. Lee, Org. Lett., 2007, 9, 33. S16