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In the snail organism, the functional metal specificity of HpCdMT and HpCuMT was contributed by metal-specific transcription programming and cell-specific expression.
To examine the metal specificity of mAP, the enzyme was dialyzed against 20 mM Tris buffer (pH 7.5) containing 5 mM EDTA.
The amino acids located in TMD V, four residues upstream from the highly conserved aspartate, seemed to be candidates of choice for determination of metal specificity of the Zn- and Fe/Zn-CDFs (histidine) or of the Mn-CDFs (aspartate).
Generally, the metal specificity of ribozymes is low, because of weaker, as compared to proteins, cation binding, so Mg2+ ions, which are always present in large amounts as stabilizers, can seemingly occupy any metal-binding site.
The metal specificity of newly identified CDF transporters can be then deduced by their classification into one of these three groups, but only an exhaustive functional characterization of CDF members will allow us to confirm this hypothesis.
Determination of the three dimensional structure of the Cd6-HpCdMT and Cu12-HpCuMT complexes is actually in progress which may unveil the detailed structural basis of the metal specificity of the two HpMT isoforms.
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This suggests that the different metal specificities of the isoforms were achieved by gene duplication and subsequent speciation by evolutionary modulation of these non-coordinating amino acid positions.
Characterization of the function, regulation and metal-specificity of metal transporters is one of the basic steps needed for the understanding of transport and accumulation of toxic metals and metalloids by plants.
This residue also plays an important role in metal specificity as mutation of Gln146 to either Leu or His in E. coli MnSOD led to impaired metalation and an inactive enzyme.
Such measurements would require an expanded repertoire of probes of different colors, increasing rigor in metal specificity, a greater number of localized probes, and careful attention to ensure no perturbation of the cellular pools.
MT metal specificity is a subject of vivid, current debate [ 16], as to a larger or lesser extent, all MTs show a degree of metal specificity in native and/or in vivo environments.
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