Inductively coupled plasma time-of-flight mass spectrometry coupled to high-performance liquid chromatography for multi-elemental speciation analysis of metalloproteins in carp cytosols

The Stoichiometric Transition from Zn6Cu1-Metallothionein to Zn7-Metallothionein Underlies the Up-regulation of Metallothionein (MT) Expression

“…The predominant species of MTs reported in cells are Zn 5 -MT and Zn 6 -MT, and they can reach up to micromolar concentrations (60). The stoichiometry of MT has been studied in purified proteins (37) or after extractions from tissues exposed to heavy metals (61,62). Recent studies have suggested that under normal physiological conditions, MTs may exist in three states of thiols as metal-bound (MT), free (T, thionein), and disulfide (T o , thionin) (63).…”

Multielement trace-element speciation in metal-biomolecules by chromatography coupled with ICP–MS

“…Unfortunately, such multielement speciation analysis has been scarce. HPLC-ICP-(TOF)MS with multielement, multi-isotopic detection was used by Goenaga Infante et al [25] for speciation analysis of metal-MT in the liver and kidneys of carp. The same detector has been used to elucidate the metal-binding patterns of the proteins expressed by mussels previously exposed to 18 different contaminating elements, in an aquarium tank, for two weeks [26].…”

Characterization of metallothionein isoforms from rabbit liver by liquid chromatography coupled to electrospray mass spectrometry

“…Both are capable of separating subisoforms on the basis of the differences between retention times in liquid chromatography or migration times in capillary electrophoresis. Some modalities of liquid chromatography, such as size exclusion chromatography, fast protein liquid chromatography, and reversed phase liquid chromatography, have been developed successfully in many MT studies [16][17][18][19][20] and applied to the analysis of MT isoforms isolated from different animal tissues [21][22][23][24]. Within the latter, reversed phase liquid chromatography is the most suitable technique for MT subisoform separations, resolving peptides whose composition differs only in one amino acid.…”