supplementary information zinc(ii) binding sites in pra1, a … · 2017-07-04 · supplementary...
TRANSCRIPT
Supplementary Information
Zinc(II) binding sites in Pra1, a zincophore from Candida albicansDorota Łoboda,a Magdalena Rowińska-Żyreka
Electronic Supplementary Material (ESI) for Dalton Transactions.This journal is © The Royal Society of Chemistry 2017
Figure S1. The alignment of Pra1 and Aspf2 sequences (Uniprot accession numbers P87020 and D3W9Z7, respectively). The two zincophores share 43% of identity.
Figure S2. Probable secondary structure of Pra1 predicted by Phyre2.
602.3
640.3
664.2
+MS,
0.0
0.5
1.0
1.5
2.0
2.5x104Intens.
600 620 640 660 680 700 720 740 760 m/z
L1+
LK1+
ZnL1+
664.2
664.7
665.2
665.7
666.2
666.7
667.2
667.7
668.2
669.2670.2
+MS,
664.2
665.2
666.2
667.2
668.2
669.2670.2
C28H38N7O8Zn,0
2000
4000
6000
8000
Intens.
0
2000
4000
6000
8000
662 664 666 668 670 672 674 m/z
ZnL1+
experimental
simulated
Figure S3. ESI-MS spectrum of Zn(II)-Ac-IEHY-NH2 at pH 6.M/L molar ratio = 1:1.
489.2
508.2520.2 527.2 546.2 565.1
+MS,
0.0
0.5
1.0
1.5
x105
Intens.
490 500 510 520 530 540 550 560 570 m/z
L2+
LK2+ZnL2+LK22+ LK32+ LK42+
520.2
520.7
521.2
521.7522.2
522.7
523.2
+MS,
520.2
520.7
521.2
521.7
522.2
522.7
523.2
C39H60N16O14Zn,0
2000
4000
6000
8000
Intens.
0
2000
4000
6000
8000
520.0 520.5 521.0 521.5 522.0 522.5 523.0 523.5 m/z
experimental
simulated
Figure S4. ESI-MS spectrum of Zn(II)-Ac-AEHARDH-NH2at pH 6.M/L molar ratio = 1:1.
589.3
608.3
620.3 627.3
+MS, 0.1-0.6min
0
1
2
3
x105Intens.
600 610 620 630 640 650 660 m/z580 590
L2+
ZnL2+
LK2+
LK22+
620.3
620.8
621.3
621.8622.3
622.8
623.3623.8 624.3
624.8625.3
+MS,
620.3
620.8
621.3
621.8622.3
622.8
623.3623.8
C58H84N18O9Zn,0.0
0.5
1.0
1.5
x104Intens.
0.0
0.5
1.0
1.5
4
621 622 623 624 625 m/z
ZnL2+
experimental
simulated
Figure S5. ESI-MS spectrum of Zn(II)-Ac-LHRFWHLK -NH2at pH 6.M/L molar ratio = 1:1.
650.3
672.3
688.3712.2
748.2
+MS,
0.0
0.5
1.0
1.5
2.0
2.5
3.0
x104Intens.
640 660 680 700 720 740 m/z
L1+
LNa1+
LK1+ZnL1+
LClO41+
+MS,
713.2
714.2
715.2
716.2
717.2718.2
C26H38N11O9Zn,0
500
1000
1500
2000
Intens.
0
500
1000
1500
2000
712 713 714 715 716 717 718 m/z
712.2
713.2
714.2
715.2
716.2
717.2
712.2
ZnL1+x104
experimental
simulated
Figure S6. ESI-MS spectrum of Zn(II)-Ac-SHQHT-NH2at pH 6.M/L molar ratio = 1:1.
486.2
492.2
504.9513.5
+MS,
0
2000
4000
6000
Intens.
480 485 490 495 500 505 510 515 520 525 m/z
L3+
[L-H2O]
LK3+ZnL3+
512.9513.2
513.5
513.9
514.2
514.5
514.9
+MS,
512.8
513.2
513.5
513.8514.2
514.5
514.8
515.2515.5
C58H81N21O21S2Zn,0
200
400
600
Intens.
0
200
400
600
800
513.0 513.5 514.0 514.5 515.0 515.5 m/z
experimental
simulated
Figure S7. ESI-MS spectrum of Zn(II)-Ac-SHCHTHADGEVHC-COOH at pH 6.M/L molar ratio = 1:1.
800.4800.6
800.8
801.1 801.3801.6
801.8
802.1
802.3
802.6
+MS,
800.5
800.8
801.0 801.3801.5
801.8
802.0
802.3
802.5802.8
803.0
C123H177N44O50S2Zn,0
100
200
300
400
500
Intens.
0
100
200
300
400
500
600
800.5 801.0 801.5 802.0 802.5 803.0 m/z
experimental
simulated
Figure S8. ESI-MS spectrum of Zn(II)-Ac-SHQHTDSNPSATTDANSHCHTHADGEVHC-COOH at pH 6. M/L molar ratio = 1:1.
Figure S9. Distribution diagram for the formation Zn(II) complex with Ac-IEHY-NH2at 25°C and I=0.1M.
2 4 6 8 100
10
20
30
40
50
60
70
80
90
100
ZnH-1L
ZnH-2LZnL
ZnHL
% fo
rmat
ion re
lative
to [Z
n2+]
pH
free Zn(II)
ZnH2L
Figure S10. Distribution diagram for the formation Zn(II) complex with Ac-AEHARDH-NH2 at 25°C and I=0.1M.
2 4 6 8 100
10
20
30
40
50
60
70
80
90
100
ZnH-2LZnH-1L
ZnHL
% fo
rmat
ion
rela
tive
to [Z
n2+]
pH
free Zn2+
ZnH2L
2 4 6 80
10
20
30
40
50
60
70
80
90
100
ZnH-2L
ZnH-1L
ZnHL
% fo
rmat
ion
rela
tive
to[Z
n2+]
pH
free Zn(II)
ZnH2L
Figure S11. Distribution diagram for the formation Zn(II) complex with Ac-LHRFWHLK-NH2at 25°C and I=0.1M.
Figure S12. Distribution diagram for the formation Zn(II) complex with Ac-SHQHT-NH2 at 25°C and I=0.1M.
2 4 6 8 100
10
20
30
40
50
60
70
80
90
100
ZnLH-2
% fo
rmat
ion
rela
tive
to [Z
n2+]
pH
free Zn ZnL
Figure S13. Distribution diagram for the formation Zn(II) complex with Ac-SHCHTHADGEVHC-COOH at 25°C and I=0.1M.
2 4 6 8 100
20
40
60
80
100
ZnL
ZnHL
ZnH2L
ZnH3L
% fo
rmat
ion
rela
tive
to [Z
n2+]
pH
free Zn2+
ZnH6L
ZnH4L
Figure S14. Distribution diagram for the formation Zn(II) complex with Ac-SHQHTDSNPSATTDANSH-CHTHADGEVHC-COOH at 25°C and I=0.1M.
2 4 6 8 100
20
40
60
80
100
ZnL
ZnHLZnH2L
% fo
rmat
ion
rela
tive
to [Z
n2+]
pH
free Zn2+
ZnH3L
Figure S15. TOCSY spectra of 3 mMAc-SHQHTDSNPSATTDANSHCHTHADGEVHC-COOH, T=298 K, in the absence (black contours) and in presence (blue contours) of 1 Zn(II) equivalent, pH 5.
Figure S16. TOCSY spectra of 3 mMAc-SHQHTDSNPSATTDANSHCHTHADGEVHC-COOH, T=298 K, in the absence (black contours) and in presence (greencontours) of 1 Zn(II) equivalent, pH 6.
Figure S17. TOCSY spectra of 3 mMAc-SHQHTDSNPSATTDANSHCHTHADGEVHC-COOH, T=298 K, in the absence (black contours) and in presence (green contours) of 1 Zn(II) equivalent, pH 6.
Figure S18. TOCSY spectra of 3 mMAc-LHRFWHLK-NH2, T=298 K, in the absence (black contours) and in presence (bright green contours) of 1 Zn(II) equivalent, pH 3.5.