Protein and metal cluster structure of the wheat metallothionein domain γ-E(c)-1: the second part of the puzzle

J Biol Inorg Chem. 2011 Jun;16(5):683-94. doi: 10.1007/s00775-011-0770-2. Epub 2011 Mar 25.

Abstract

Metallothioneins (MTs) are small cysteine-rich proteins coordinating various transition metal ions, including Zn(II), Cd(II), and Cu(I). MTs are ubiquitously present in all phyla, indicating a successful molecular concept for metal ion binding in all organisms. The plant MT E(c)-1 from Triticum aestivum, common bread wheat, is a Zn(II)-binding protein that comprises two domains and binds up to six metal ions. The structure of the C-terminal four metal ion binding β(E) domain was recently described. Here we present the structure of the N-terminal second domain, γ-E(c)-1, determined by NMR spectroscopy. The γ-E(c)-1 domain enfolds an M (2) (II) Cys(6) cluster and was characterized as part of the full-length Zn(6)E(c)-1 protein as well as in the form of the separately expressed domain, both in the Zn(II)-containing isoform and the Cd(II)-containing isoform. Extended X-ray absorption fine structure analysis of Zn(2)γ-E(c)-1 clearly shows the presence of a ZnS(4) coordination sphere with average Zn-S distances of 2.33 Å. (113)Cd NMR experiments were used to identify the M(II)-Cys connectivity pattern, and revealed two putative metal cluster conformations. In addition, the general metal ion coordination abilities of γ-E(c)-1 were probed with Cd(II) binding experiments as well as by pH titrations of the Zn(II) and Cd(II) forms, the latter suggesting an interaction of the γ domain and the β(E) domain within the full-length protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cadmium / metabolism
  • Metallothionein / chemistry*
  • Metallothionein / metabolism
  • Metals / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular
  • Plant Proteins / chemistry*
  • Plant Proteins / metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Triticum / chemistry*
  • Triticum / metabolism
  • Zinc / metabolism

Substances

  • Metals
  • Plant Proteins
  • Cadmium
  • Metallothionein
  • Zinc