The Protein Journal

, Volume 24, Issue 6, pp 327–336 | Cite as

The Properties of the Metal–Thiolate Clusters in Recombinant Mouse Metallothionein-4

  • Bin Cai
  • Qi Zheng
  • Zhong-Xian Huang

Metallothioneins (MTs) are metal-binding proteins with low molecular weight and conservative cysteine residues. Metallothionein-4 (MT-4), one of MT isoforms, is first reported to be distributed in a tissue-specific manner, mainly in stratified squamous epithelia. Here, we compare the properties of metal–thiolate clusters in MT-4 to those in MT-1 and MT-3, including the stabilities toward both pH change and EDTA, as well as the exposure of thiolates to solvent. The metal–thiolate clusters in MT-3 show different property and activity to the reactions compared with MT-4 and MT-1. The structure of metal–thiolate clusters in MT-4 is similar to that of MT-1 from the UV and CD spectra. During pH titration and DTNB reaction, MT-4 and MT-1 exhibit comparable behavior. But while reacting with EDTA, the metal–thiolate clusters in MT-4 are more stable than those of MT-1. We suppose the negative charge of the β-domain of MT-4 prevents the EDTA attack to MT-4.


DTNB EDTA metallothionein pH titration 



Circular dichroism


5,5′-dithiobis-2-nitrobenzoic acid


Ethylenediamine tetraacetic acid


Electrospray ionization-mass spectroscopy


glutathione sulfur transferase


Ligand metal charge transfer




Ultraviolet visible


zinc-transport proteins.


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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Chemical Biology Lab, Department of ChemistryFudan UniversityShanghaiChina

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