The two Caenorhabditis elegans metallothioneins (CeMT-1 and CeMT-2) discriminate between essential zinc and toxic cadmium.
The nematode Caenorhabditis elegans expresses two metallothioneins (MTs), CeMT-1 and CeMT-2, that are believed to be key players in the protection against metal toxicity. In this study, both isoforms were expressed in vitro in the presence of either Zn(II) or Cd(II). Metal binding stoichiometries and affinities were determined by ESI-MS and NMR, respectively. Both isoforms had equal zinc binding ability, but differed in their cadmium binding behaviour, with higher affinity found for CeMT-2. In addition, wild-type C. elegans, single MT knockouts and a double MT knockout allele were exposed to zinc (340 microm) or cadmium (25 microm) to investigate effects in vivo. Zinc levels were significantly increased in all knockout strains, but were most pronounced in the CeMT-1 knockout, mtl-1 (tm1770), while cadmium accumulation was highest in the CeMT-2 knockout, mtl-2 (gk125) and the double knockout mtl-1;mtl-2 (zs1). In addition, metal speciation was assessed by X-ray absorption fine-structure spectroscopy. This showed that O-donating, probably phosphate-rich, ligands play a dominant role in maintaining the physiological concentration of zinc, independently of metallothionein status. In contrast, cadmium was shown to coordinate with thiol groups, and the cadmium speciation of the wild-type and the CeMT-2 knockout strain was distinctly different to the CeMT-1 and double knockouts. Taken together, and supported by a simple model calculation, these findings show for the first time that the two MT isoforms have differential affinities towards Cd(II) and Zn(II) at a cellular level, and this is reflected at the protein level. This suggests that the two MT isoforms have distinct in vivo roles.
Zeitoun-Ghandour
Sukaina
S
School of Biomedical & Health Sciences, King's College London, London, UK.
Charnock
John M
JM
Hodson
Mark E
ME
Leszczyszyn
Oksana I
OI
Blindauer
Claudia A
CA
Stürzenbaum
Stephen R
SR
D000818:Animals; D002104:Cadmium / Q000378:metabolism / Q000633:toxicity*; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000187:drug effects / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008668:Metallothionein / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D056928:X-Ray Absorption Spectroscopy; D015032:Zinc / Q000378:metabolism*
D029742:Caenorhabditis elegans Proteins; D015801:Helminth Proteins; D020033:Protein Isoforms; D002104:Cadmium; D008668:Metallothionein; D015032:Zinc