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Locating Pb2+ and Zn2+ in Zinc Finger-Like Peptides Using Mass Spectrometry

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

The binding preferences of Pb2+and Zn2+ in doubly charged complexes with zinc finger-like 12-residue peptides (Pep), [Mn(Pep-2(n-1)H)]2+ have been explored using tandem mass spectrometry. The peptides were synthesized strategically by blocking the N-terminus with an acetyl group and with four cysteine and/or histidine residues in positions 2, 5, 8, and 11, arranged in different motifs: CCHH, CHCH, and CCCC. The MS2 spectra of the Pb2+ and Zn2+ complexes show multiple losses of water and a single methane loss and these provide a sensitive method for locating the metal dication and so elucidating its coordination. The elimination of a methane molecule indicated the position of the metal at the Cys2 residue. Whereas lead was observed to preferentially bind to cysteine residues, zinc was found to primarily bind to histidine residues and secondarily to cysteine residues. Preferential binding of lead to cysteine is preserved in the complexes with more than one Pb2+. Key to the mechanism of the loss of water and methane is the metal dication withdrawing electrons from the proximal amidic nitrogen. This acidic nitrogen loses its hydrogen to an amidic oxygen situated four atoms away leading to formation of a five-member ring and the elimination of water.

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Acknowledgments

The authors greatly appreciate continued funding from the Natural Sciences and Engineering Research Council of Canada. As holder of a Canada Research Chair in Physical Chemistry, D.K.B. thanks the contributions of the Canada Research Chair Program to this research.

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Authors and Affiliations

  1. Department of Chemistry, Centre for Research in Mass Spectrometry, York University, Toronto, Ontario, M3J 1P3, Canada

    Laura Banu, Voislav Blagojevic & Diethard K. Bohme

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  1. Laura Banu
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  2. Voislav Blagojevic
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  3. Diethard K. Bohme
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Correspondence to Diethard K. Bohme.

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Banu, L., Blagojevic, V. & Bohme, D.K. Locating Pb2+ and Zn2+ in Zinc Finger-Like Peptides Using Mass Spectrometry. J. Am. Soc. Mass Spectrom. 24, 1534–1542 (2013). https://doi.org/10.1007/s13361-013-0682-1

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