title: Dual nanomolar and picomolar Zn(II) binding properties of metallothionein.
abstract: Each of the seven Zn(II) ions in the Zn(3)S(9) and Zn(4)S(11) clusters of human metallothionein is in a tetrathiolate coordination environment. Yet analysis of Zn(II) association with thionein, the apoprotein, and analysis of Zn(II) dissociation from metallothionein using the fluorescent chelating agents FluoZin-3 and RhodZin-3 reveal at least three classes of sites with affinities that differ by 4 orders of magnitude. Four Zn(II) ions are bound with an apparent average log K of 11.8, and with the methods employed, their binding is indistinguishable. This binding property makes thionein a strong chelating agent. One Zn(II) ion is relatively weakly bound, with a log K of 7.7, making metallothionein a zinc donor in the absence of thionein. The binding data demonstrate that Zn(II) binds with at least four species: Zn(4)T, Zn(5)T, Zn(6)T, and Zn(7)T. Zn(5)T and Zn(6)T bind Zn(II) with a log K of approximately 10 and are the predominant species at micromolar concentrations of metallothionein in cells. Central to the function of the protein is the reactivity of its cysteine side chains in the absence and presence of Zn(II). Chelating agents, such as physiological ligands with moderate affinities for Zn(II), cause dissociation of Zn(II) ions from metallothionein at pH 7.4 (Zn(7)T <==> Zn(7-n)T + nZn(2+)), thereby affecting the reactivity of its thiols. Thus, the rate of thiol oxidation increases in the presence of Zn(II) acceptors but decreases if more free Zn(II) becomes available. Thionein is such an acceptor. It regulates the reactivity and availability of free Zn(II) from metallothionein. At thionein/metallothionein ratios > 0.75, free Zn(II) ions are below a pZn (-log[Zn(2+)](free)) of 11.8, and at ratios < 0.75, relatively large fluctuations of free Zn(II) ions are possible (pZn between 7 and 11). These chemical characteristics match cellular requirements for Zn(II) and suggest how the molecular structures and redox chemistries of metallothionein and thionein determine Zn(II) availability for biological processes.
authors: Department of Preventive Medicine and Community Health, The University of Texas Medical Branch, Galveston, Texas 77555, USA.
authors: Artur
authors: A
authors: Krezel
authors: Wolfgang
authors: W
authors: Maret
mesh_terms: D000595:Amino Acid Sequence; D002413:Cations, Divalent; D004880:Ergothioneine / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008668:Metallothionein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D008024:Ligands; D011994:Recombinant Proteins; D008668:Metallothionein; D004880:Ergothioneine; D015032:Zinc