The CdII-binding abilities of recombinant Quercus suber metallothionein, QsMT: bridging the gap between phytochelatins and metallothioneins
J. Domènech, R. Orihuela, G. Mir, M. Molinas, S. Atrian, M. Capdevila
SUPPLEMENTARY MATERIAL
Tables
Table S1 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-rQsMT with CdCl2 at pH 7 as a function of the number of CdII or S2- eq added.
Table S2 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-N25-C18 with CdCl2 at pH 7 as a function of the number of CdII eq added.
Table S3 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-N25 with CdCl2 at pH 7 as a function of the number of CdII eq added.
Table S4 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-C18 with CdCl2 at pH 7 as a function of the number of CdII eq added.
Table S5 Distribution of the metal complexes present in solution, according to ESI-MS data, during the co-titration of ZnII-C18 and ZnII-N25 with CdCl2 at pH 7 as a function of the number of CdII eq added.
Figures
Fig. S1 CD spectra corresponding to the acidification of a 20 µM solution of CdII-rQsMT(1) from pH 7.0 to pH 1.0.
Fig. S2 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-N25-C18 with CdII at pH 7.0 followed by the addition of several Na2S eq.
Fig. S3 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first two rows), reneutralization (third row) and addition of several Na2S eq (fourth row) of a 20 µM solution of CdII-N25-C18.
Fig. S4 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-N25 with CdII at pH 7.0 followed of the addition of several Na2S eq.
Fig. S5 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-C18 with CdII at pH 7.0 followed by the addition of several Na2S eq.
Fig. S6 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first two rows), reneutralization (third and fourth row) and addition of several Na2S eq (fifth row) of a 20 µM solution of CdII-N25.
Fig. S7 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first three rows), reneutralization (fourth row) and addition of several Na2S eq (fifth row) of a 20 µM solution of CdII-C18.
CdII and/or S2- eq added to ZnII-rQsMT4 CdII / 7 CdII / 7 CdII + 3 S2- * / 7 CdII + 4 S2- *
Zn1Cd3-QsMT / /
Zn1Cd4-QsMT / /
Zn1Cd5-QsMT /
Cd3-QsMT / /
Cd3S2-QsMT /
Cd4-QsMT / / /
Cd5-QsMT / / x / x
Cd6-QsMT / x /
Cd6S6-QsMT / x
Cd7S3-QsMT / x
Cd7S9-QsMT /
Table S1 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnIIrQsMT with CdCl2 at pH 7 as a function of the number of CdII or S2- eq added.
(Code: denotes the major species; x species of intermediate abundance and , the minor species)
* The high signal-to-noise ratio due to the numerous Na-adducts formed hampers a more detailed interpretation of the MS data.
CdII eq added to ZnII-N25-C185 CdII / 6 CdII / 10 CdII
Cd5-N25-C18 / / /
Cd4-N25-C18 / x / x / x
Cd6-N25-C18 / / x / x
Cd5S1-N25-C18 / x
Table S2 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-N25-C18 with CdCl2 at pH 7 as a function of the number of CdII eq added.
(Code: denotes the major species; x species of intermediate abundance and , the minor species)
Note: ESI-MS data corresponding to the addition of sulfide anions are not included due to the difficulties encountered when interpreting the spectra because of the Na-adducts detected.
CdII eq added to ZnII-N252 CdII / 4 CdII / 8CdII / 10 CdII
Zn1Cd2-N25 / x
Zn1Cd3-N25 / /
Cd2-N25 / / / /
Cd3-N25 / / / /
Cd3S2-N25 / / x
Cd4-N25 /
Table S3 Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-N25 with CdCl2 at pH 7 as a function of the number of CdII eq added.
(Code: denotes the major species; x species of intermediate abundance and , the minor species)
Note: ESI-MS data corresponding to the addition of sulfide anions are not included due to the difficulties encountered when interpreting the spectra because of the Na-adducts detected.
CdII eq added to ZnII-C181 CdII / 2 CdII / 3 CdII
Zn2Cd2-(C18)2 / Zn1Cd1-C18 / / x
Cd4-(C18)2 / Cd2-C18 / x / /
Cd2-(C18)2 / x
Zn1Cd2-(C18)2 /
Cd4S1-(C18)2 /
Cd3Zn3-(C18)2 / x
Table S4Distribution of the metal complexes present in solution, according to ESI-MS data, during the titration of ZnII-C18 with CdCl2 at pH 7 as a function of the number of CdII eq added.
(Code: denotes the major species; x species of intermediate abundance and , the minor species)
Note: ESI-MS data corresponding to the addition of sulfide anions are not included due to the difficulties encountered when interpreting the spectra because of the Na-adducts detected.
CdII eq added to ZnII-N25 + ZnII-C182 CdII / 4 CdII / 6 CdII
Zn1Cd1-C18 /Zn2Cd2-(C18)2 / / x
Cd1-C18 /Cd2-(C18)2 / x /
Cd4-(C18)2 / / x /
Cd2-C18
Zn2Cd5-(N25)2 / Cd5S1-(N25)2 / /
Cd3-(N25) / /
Cd4S1-(N25)2 / x
Cd5-(N25-C18) /
Table S5Distribution of the metal complexes present in solution, according to ESI-MS data, during the co-titration of ZnII-C18 and ZnII-N25 with CdCl2 at pH 7 as a function of the number of CdII eq added.
(Code: denotes the major species; x species of intermediate abundance and , the minor species
Note: ESI-MS data corresponding to the addition of sulfide anions are not included due to the difficulties encountered when interpreting the spectra because of the Na-adducts detected.
Fig. S1CD spectra corresponding to the acidification of a 20 µM solution of CdII-rQsMT(1) from pH 7.0 to pH 1.0.
ABC
Fig. S2CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-N25-C18 with CdII at pH 7.0 followed by the addition of several Na2S eq. The CdII or S2- to MT molar ratio are indicated within each frame.
ABC
Fig. S3 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first two rows), reneutralization (third row) and addition of several Na2S eq (fourth row) of a 20 µM solution of CdII-N25-C18. The pH and S2- to MT molar ratio are indicated within each frame.
ABC
Fig. S4CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-N25 with CdII at pH 7.0 followed of the addition of several Na2S eq. The CdII or S2- to MT molar ratio are indicated within each frame.
ABC
Fig. S5 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the titration of a 20 µM solution of ZnII-C18 with CdII at pH 7.0 followed by the addition of several Na2S eq. The CdII or S2- to MT molar ratio are indicated within each frame.
ABC
Fig. S6 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first two rows), reneutralization (third and fourth row) and addition of several Na2S eq (fifth row) of a 20 µM solution of CdII-N25. The pH and S2- to MT molar ratio are indicated within each frame.
ABC
Fig. S7 CD (A), UV-vis (B) and UV-vis difference (C) spectra corresponding to the acidification (first three rows), reneutralization (fourth row) and addition of several Na2S eq (fifth row) of a 20 µM solution of CdII-C18. The pH and S2- to MT molar ratio are indicated within each frame.
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