title: Zinc transfer from the embryo-specific metallothionein E(C) from wheat: a case study.
abstract: The direct observation of binding and release of spectroscopically silent metal ions such as Zn(2+) and Ca(2+) by proteins has been challenging before the advent of native electrospray ionisation mass spectrometry. This report highlights the powerful capability of ESI-MS to provide insight into metalloprotein speciation that is independent of any spectroscopic property. Using the zinc-binding plant metallothionein E(C) from wheat as a study case, we show that ESI-MS is unique amongst other techniques in capturing intermediary metallospecies that evolve during the course of metal transfer to the chelator EDTA, as a model reaction to mimic the biological function of the protein as a zinc donor. Zinc release from the two-domain protein E(C) appears to be extremely rapid and non-cooperative, and progresses with loss of one zinc ion from the fully loaded Zn(6) species, and a transient build-up of Zn(5) and Zn(4) species, which further react to give species with 0-3 zinc ions bound. (1)H NMR data has provided further insights into the different behaviour of the two domains upon metal depletion.
authors.affiliation: Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Oksana I
authors.initials: OI
authors.lastname: Leszczyszyn
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Blindauer
mesh_terms: D000595:Amino Acid Sequence; D004492:Edetic Acid / Q000737:chemistry; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008668:Metallothionein / Q000737:chemistry*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D014908:Triticum / Q000378:metabolism*; D015032:Zinc / Q000737:chemistry*
chemical_list: D008668:Metallothionein; D004492:Edetic Acid; D015032:Zinc