Copper Environment in Artificial Metalloproteins Probed by Electron Paramagnetic Resonance Spectroscopy

Marco Flores, Tien L. Olson, Dong Wang, Selvakumar Edwardraja, Sandip Shinde, Jo Ann C. Williams, Giovanna Ghirlanda, James P. Allen

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The design of binding sites for divalent metals in artificial proteins is a productive platform for examining the characteristics of metal-ligand interactions. In this report, we investigate the spectroscopic properties of small peptides and four-helix bundles that bind Cu(II). Three small peptides, consisting of 15 amino acid residues, were designed to have two arms, each containing a metal-binding site comprised of different combinations of imidazole and carboxylate side chains. Two four-helix bundles each had a binding site for a central dinuclear metal cofactor, with one design incorporating additional potential metal ligands at two identical sites. The small peptides displayed pH-dependent, metal-induced changes in the circular dichroism spectra, consistent with large changes in the secondary structure upon metal binding, while the spectra of the four-helix bundles showed a predominant α-helix content but only small structural changes upon metal binding. Electron paramagnetic resonance spectra were measured at X-band revealing classic Cu(II) axial patterns with hyperfine coupling peaks for the small peptides and four-helix bundles exhibiting a range of values that were related to the specific chemical natures of the ligands. The variety of electronic structures allow us to define the distinctive environment of each metal-binding site in these artificial systems, including the designed additional binding sites in one of the four-helix bundles.

Original languageEnglish
Pages (from-to)13825-13833
Number of pages9
JournalJournal of Physical Chemistry B
Volume119
Issue number43
DOIs
Publication statusPublished - Oct 29 2015

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ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Surfaces, Coatings and Films
  • Materials Chemistry

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