Conformational study of an artificial metal-dependent regulation site for use in designer proteins

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Conformational study of an artificial metal-dependent regulation site for use in designer proteins. / Oheix, Emmanuel; Spencer, Neil; Gethings, Lee A.; Peacock, Anna F. A.

In: Zeitschrift fur Anorganische und Allgemeine Chemie, Vol. 639, No. 8-9, 07.2013, p. 1370-1383.

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@article{ab0721f0a0364acebf8a610ef6a97c2f,
title = "Conformational study of an artificial metal-dependent regulation site for use in designer proteins",
abstract = "This report describes the dimerisation of glutathione, and by extension, other cysteine-containing peptides or protein fragments, with a 5, 5'-disubstituted-2, 2'-bipyridine or 6, 6''-disubstituted-2, 2':6',2''-terpyridine unit. The resulting bipy-GS2 and terpy-GS2 were investigated as potential metal ion dependent switches in aqueous solution, and were found to predominantly adopt the transo{\"i}d conformation at physiological pH. Metal complexation with CuII and ZnII at this pH has been studied by UV/Vis, CD, NMR and ion-mobility mass spectrometry. ZnII titrations are consistent with the formation of a 1:1 ZnII:terpy-GS2 complex at pH 7.4, but bipy-GS2 was shown to form both 1:1 and 1:2 complexes with the former being predominant under dilute micromolar conditions. Formation constants for the resulting 1:1 complexes were determined to be log KM 6.86 (bipy-GS2) and 6.22 (terpy-GS2), consistent with a higher affinity for the unconstrained bipyridine, compared to the strained terpyridine. CuII coordination involves the initial formation of 1:1 complexes, followed by 1.5Cu:1bipy-GS2 and 2Cu:1terpy-GS2 complexes at micromolar concentrations. Binding constants for formation of the 1:1 complexes (log KM 12.5 (bipy-GS2); 8.04 and 7.14 (terpy-GS2)) indicate a higher affinity for CuII than ZnII. Finally, ion-mobility MS studies detected the free ligands in their protonated form, and were consistent with the formation of two different Cu adducts with different conformations in the gas-phase. We illustrate that the bipyridine and terpyridine dimerisation units can behave like conformational switches in response to Cu/Zn complexation, and propose that in future these can be employed in synthetic biology with larger peptide or protein fragments, to control large scale folding and related biological function.",
keywords = "Bioinorganic chemistry, Metalloswitches, Bipyridine, Terpyridine, Peptides",
author = "Emmanuel Oheix and Neil Spencer and Gethings, {Lee A.} and Peacock, {Anna F. A.}",
year = "2013",
month = "7",
doi = "10.1002/zaac.201300131",
language = "English",
volume = "639",
pages = "1370--1383",
journal = "Zeitschrift fur Anorganische und Allgemeine Chemie",
issn = "0044-2313",
publisher = "Wiley-VCH Verlag",
number = "8-9",

}

RIS

TY - JOUR

T1 - Conformational study of an artificial metal-dependent regulation site for use in designer proteins

AU - Oheix, Emmanuel

AU - Spencer, Neil

AU - Gethings, Lee A.

AU - Peacock, Anna F. A.

PY - 2013/7

Y1 - 2013/7

N2 - This report describes the dimerisation of glutathione, and by extension, other cysteine-containing peptides or protein fragments, with a 5, 5'-disubstituted-2, 2'-bipyridine or 6, 6''-disubstituted-2, 2':6',2''-terpyridine unit. The resulting bipy-GS2 and terpy-GS2 were investigated as potential metal ion dependent switches in aqueous solution, and were found to predominantly adopt the transoïd conformation at physiological pH. Metal complexation with CuII and ZnII at this pH has been studied by UV/Vis, CD, NMR and ion-mobility mass spectrometry. ZnII titrations are consistent with the formation of a 1:1 ZnII:terpy-GS2 complex at pH 7.4, but bipy-GS2 was shown to form both 1:1 and 1:2 complexes with the former being predominant under dilute micromolar conditions. Formation constants for the resulting 1:1 complexes were determined to be log KM 6.86 (bipy-GS2) and 6.22 (terpy-GS2), consistent with a higher affinity for the unconstrained bipyridine, compared to the strained terpyridine. CuII coordination involves the initial formation of 1:1 complexes, followed by 1.5Cu:1bipy-GS2 and 2Cu:1terpy-GS2 complexes at micromolar concentrations. Binding constants for formation of the 1:1 complexes (log KM 12.5 (bipy-GS2); 8.04 and 7.14 (terpy-GS2)) indicate a higher affinity for CuII than ZnII. Finally, ion-mobility MS studies detected the free ligands in their protonated form, and were consistent with the formation of two different Cu adducts with different conformations in the gas-phase. We illustrate that the bipyridine and terpyridine dimerisation units can behave like conformational switches in response to Cu/Zn complexation, and propose that in future these can be employed in synthetic biology with larger peptide or protein fragments, to control large scale folding and related biological function.

AB - This report describes the dimerisation of glutathione, and by extension, other cysteine-containing peptides or protein fragments, with a 5, 5'-disubstituted-2, 2'-bipyridine or 6, 6''-disubstituted-2, 2':6',2''-terpyridine unit. The resulting bipy-GS2 and terpy-GS2 were investigated as potential metal ion dependent switches in aqueous solution, and were found to predominantly adopt the transoïd conformation at physiological pH. Metal complexation with CuII and ZnII at this pH has been studied by UV/Vis, CD, NMR and ion-mobility mass spectrometry. ZnII titrations are consistent with the formation of a 1:1 ZnII:terpy-GS2 complex at pH 7.4, but bipy-GS2 was shown to form both 1:1 and 1:2 complexes with the former being predominant under dilute micromolar conditions. Formation constants for the resulting 1:1 complexes were determined to be log KM 6.86 (bipy-GS2) and 6.22 (terpy-GS2), consistent with a higher affinity for the unconstrained bipyridine, compared to the strained terpyridine. CuII coordination involves the initial formation of 1:1 complexes, followed by 1.5Cu:1bipy-GS2 and 2Cu:1terpy-GS2 complexes at micromolar concentrations. Binding constants for formation of the 1:1 complexes (log KM 12.5 (bipy-GS2); 8.04 and 7.14 (terpy-GS2)) indicate a higher affinity for CuII than ZnII. Finally, ion-mobility MS studies detected the free ligands in their protonated form, and were consistent with the formation of two different Cu adducts with different conformations in the gas-phase. We illustrate that the bipyridine and terpyridine dimerisation units can behave like conformational switches in response to Cu/Zn complexation, and propose that in future these can be employed in synthetic biology with larger peptide or protein fragments, to control large scale folding and related biological function.

KW - Bioinorganic chemistry

KW - Metalloswitches

KW - Bipyridine

KW - Terpyridine

KW - Peptides

U2 - 10.1002/zaac.201300131

DO - 10.1002/zaac.201300131

M3 - Article

VL - 639

SP - 1370

EP - 1383

JO - Zeitschrift fur Anorganische und Allgemeine Chemie

JF - Zeitschrift fur Anorganische und Allgemeine Chemie

SN - 0044-2313

IS - 8-9

ER -