The two distinctive metal ion binding domains of the wheat metallothionein Ec-1.
Metallothioneins are small cysteine-rich proteins believed to play a role, among others, in the homeostasis of essential metal ions such as Zn(II) and Cu(I). Recently, we could show that wheat E(c)-1 is coordinating its six Zn(II) ions in form of metal-thiolate clusters analogously to the vertebrate metallothioneins. Specifically, two Zn(II) ions are bound in the N-terminal and four in the C-terminal domain. In the following, we will present evidence for the relative independence of the two domains from each other with respect to their metal ion binding abilities, and uncover three intriguing peculiarities of the protein. Firstly, one Zn(II) ion of the N-terminal domain is relative resistant to complete replacement with Cd(II) indicating the presence of a Zn(II)-binding site with increased stability. Secondly, the C-terminal domain is able to coordinate an additional fifth metal ion, though with reduced affinity, which went undetected so far. Finally, reconstitution of apoE(c)-1 with an excess of Zn(II) shows a certain amount of sub-stoichiometrically metal-loaded species. The possible relevance of these finding for the proposed biological functions of wheat E(c)-1 will be discussed. In addition, extended X-ray absorption fine structure (EXAFS) measurements on both, the full-length and the truncated protein, provide final evidence for His participation in metal ion binding.
Peroza
Estevão A
EA
Institute of Inorganic Chemistry, University of Zürich, 8057 Zürich, Switzerland.
Kaabi
Ali Al
AA
Meyer-Klaucke
Wolfram
W
Wellenreuther
Gerd
G
Freisinger
Eva
E
D001059:Apoproteins / Q000378:metabolism*; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism*; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary / Q000235:genetics; D014908:Triticum / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*
D001059:Apoproteins; D002104:Cadmium; D008668:Metallothionein; D015032:Zinc