The beta(E)-domain of wheat E(c)-1 metallothionein: a metal-binding domain with a distinctive structure.
Metallothioneins (MTs) are ubiquitous cysteine-rich proteins with a high affinity for divalent metal ions such as Zn(II), Cu(I), and Cd(II) that are involved in metal ion homeostasis and detoxification, as well as protection against reactive oxygen species. Here we show the NMR solution structure of the beta(E)-domain of the early cysteine-labeled protein (E(c)-1) from wheat (beta(E)-E(c)-1), which represents the first three-dimensional structure of a plant MT. The beta(E)-domain comprises the 51 C-terminal residues of E(c)-1 and exhibits a distinctive unprecedented structure with two separate metal-binding centers, a mononuclear Zn(II) binding site constituted by two cysteine and two highly conserved histidine residues as found in certain zinc-finger motifs, and a cluster formed by three Zn(II) ions coordinated by nine Cys residues that resembles the cluster in the beta-domain of vertebrate MTs. Cys-metal ion connectivities were determined by exhaustive structure calculations for all 7560 possible configurations of the three-metal cluster. Backbone dynamics investigated by (15)N relaxation experiments support the results of the structure determination in that beta(E)-E(c)-1 is a rigidly folded polypeptide. To further investigate the influence of metal ion binding on the stability of the structure, we replaced Zn(II) with Cd(II) ions and examined the effects of metal ion release on incubation with a metal ion chelator.
Institute of Inorganic Chemistry, University of Zürich, Zürich, Switzerland.
Estevão A
EA
Peroza
Roland
R
Schmucki
Peter
P
Güntert
Eva
E
Freisinger
Oliver
O
Zerbe
D000595:Amino Acid Sequence; D001665:Binding Sites; D008668:Metallothionein / Q000737:chemistry / Q000378:metabolism*; D008670:Metals / Q000378:metabolism*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet; D014908:Triticum / Q000737:chemistry*
D008670:Metals; D008668:Metallothionein