The application of ¹⁹⁹Hg NMR and ^199mHg perturbed angular correlation (PAC) spectroscopy to define the biological chemistry of Hg^II: a case study with designed two- and three-stranded coiled coils

TY - JOUR

T1 - The application of 199Hg NMR and 199mHg perturbed angular correlation (PAC) spectroscopy to define the biological chemistry of HgII

T2 - a case study with designed two- and three-stranded coiled coils

AU - Iranzo, Olga

AU - Thulstrup, Peter Waaben

AU - Ryu, Seung-baek

AU - Hemmingsen, Lars Bo Stegeager

AU - Pecoraro, Vincent L.

PY - 2007

Y1 - 2007

N2 - The use of de novo designed peptides is a powerful strategy to elucidate HgII-protein interactions and to gain insight into the chemistry of HgII in biological systems. Cysteine derivatives of the designed -helical peptides of the TRI family [Ac-G-(LaKbAcLdEeEfKg)4-G-NH2] bind HgII at high pH values and at peptide/HgII ratios of 3:1 with an unusual trigonal thiolate coordination mode. The resulting HgII complexes are good water-soluble models for HgII binding to the protein MerR. We have carried out a parallel study using 199Hg NMR and 199mHg perturbed angular correlation (PAC) spectroscopy to characterize the distinct species that are generated under different pH conditions and peptide TRI L9C/HgII ratios. These studies prove for the first time the formation of [Hg{(TRI L9C)2-(TRI L9CH)}], a dithiolate-HgII complex in the hydrophobic interior of the three-stranded coiled coil (TRI L9C)3. 199Hg NMR and 199mHg PAC data demonstrate that this dithiolate-HgII complex is different from the dithiolate [Hg(TRI L9C)2], and that the presence of third -helix, containing a protonated cysteine, breaks the symmetry of the coordination environment present in the complex [Hg(TRI L9C)2]. As the pH is raised, the deprotonation of this third cysteine generates the trigonal thiolate-HgII complex Hg(TRI L9C)3- on a timescale that is slower than the NMR timescale (0.01-10 ms). The formation of the species [Hg{(TRI L9C)2(TRI L9CH)}] is the result of a compromise between the high affinity of HgII to form dithiolate complexes and the preference of the peptide to form a three-stranded coiled coil.

AB - The use of de novo designed peptides is a powerful strategy to elucidate HgII-protein interactions and to gain insight into the chemistry of HgII in biological systems. Cysteine derivatives of the designed -helical peptides of the TRI family [Ac-G-(LaKbAcLdEeEfKg)4-G-NH2] bind HgII at high pH values and at peptide/HgII ratios of 3:1 with an unusual trigonal thiolate coordination mode. The resulting HgII complexes are good water-soluble models for HgII binding to the protein MerR. We have carried out a parallel study using 199Hg NMR and 199mHg perturbed angular correlation (PAC) spectroscopy to characterize the distinct species that are generated under different pH conditions and peptide TRI L9C/HgII ratios. These studies prove for the first time the formation of [Hg{(TRI L9C)2-(TRI L9CH)}], a dithiolate-HgII complex in the hydrophobic interior of the three-stranded coiled coil (TRI L9C)3. 199Hg NMR and 199mHg PAC data demonstrate that this dithiolate-HgII complex is different from the dithiolate [Hg(TRI L9C)2], and that the presence of third -helix, containing a protonated cysteine, breaks the symmetry of the coordination environment present in the complex [Hg(TRI L9C)2]. As the pH is raised, the deprotonation of this third cysteine generates the trigonal thiolate-HgII complex Hg(TRI L9C)3- on a timescale that is slower than the NMR timescale (0.01-10 ms). The formation of the species [Hg{(TRI L9C)2(TRI L9CH)}] is the result of a compromise between the high affinity of HgII to form dithiolate complexes and the preference of the peptide to form a three-stranded coiled coil.

U2 - 10.1002/chem.200701208

DO - 10.1002/chem.200701208

M3 - Journal article

C2 - 17960740

SN - 0947-6539

VL - 13

SP - 9178

EP - 9190

JO - Chemistry: A European Journal

JF - Chemistry: A European Journal

IS - 33

ER -