The metal binding abilities of Megathura crenulata metallothionein (McMT) in the frame of gastropoda MTs

Metallothioneins (MTs) are proteins that play a major role in metal homeostasis and/or detoxification in all kind of organisms. The MT gene/protein system of gastropod molluscs provides an invaluable model to study the diversification mechanisms that have enabled MTs to achieve metal-binding specificity through evolution. Most pulmonate gastropods, particularly terrestrial snails, harbor three paralogous isogenes encoding three MT isoforms with different metal binding preferences: the highly specific CdMT and CuMT isoforms, for cadmium and copper respectively, and the unspecific Cd/CuMT isoform. Megathura crenulata is a non-pulmonate gastropod in which only one MT isogene has so far been reported. In order to elucidate the metal binding character of the corresponding peptide (McMT), it has been recombinantly synthesized in the presence of Cd(2+), Zn(2+) or Cu(2+), and the corresponding metal complexes have been analyzed using electrospray mass spectrometry, and CD and UV-visible spectroscopy. The metal-binding traits exhibited by McMT revealed that it is an unspecific MT, similarly to the pulmonate Cd/CuMT isoforms. This is in full concordance with the protein sequence distance analysis in relation to other gastropod MTs.