Structural changes in metallothionein isoforms revealed by capillary electrophoresis and Brdicka reaction
Language English Country Germany Media print
Document type Journal Article
- MeSH
- Biochemical Phenomena MeSH
- Chemical Precipitation MeSH
- Models, Chemical * MeSH
- Protein Denaturation MeSH
- Electrophoresis, Capillary methods MeSH
- Rabbits MeSH
- Linear Models MeSH
- Metallothionein chemistry metabolism MeSH
- Urea chemistry MeSH
- Molecular Sequence Data MeSH
- Biomarkers, Tumor analysis chemistry MeSH
- Protein Isoforms MeSH
- Amino Acid Sequence MeSH
- Sequence Alignment MeSH
- Sensitivity and Specificity MeSH
- Spectrophotometry, Ultraviolet MeSH
- Hot Temperature MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Metallothionein MeSH
- Urea MeSH
- Biomarkers, Tumor MeSH
- Protein Isoforms MeSH
Metallothionein (MT) as a potential cancer marker is at the center of interest and its properties, functions and behavior under various conditions is intensively studied. In the present study, two major mammalian MT isoforms (MT-1 and MT-2) were separated using capillary electrophoresis (CE) coupled with UV detector in order to describe their basic behavior. Under the optimized conditions, the separation of both isoforms was enabled as well as estimation of detection limits as subunits and units of ng per μL for MT-2 and MT-1, respectively. Further, the effects of thermal treatment and the presence of denaturing agent such as urea on MT-1 and MT-2 isoforms were studied by CE-UV. Thermal treatment caused an increase in the signals of both isoforms. A new parameter called precipitation rate has been defined based on this finding. This parameter can be expressed as a slope of the linear regression of the time dependency curve recalculated on the MT concentration. The thermal precipitation rate for MT-1 and MT-2 was determined as 1.1 and 0.9 ng of MT/min, respectively. The chemical precipitation rate calculated from the linear regression for both isoforms provided the same value of 0.25 ng of MT/min. The results were confirmed by manual spectrometric measurements and by differential pulse voltammetry Brdicka reaction. Based on these results, a model of MT behavior under the conditions studied was suggested.
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Fluorescence-tagged metallothionein with CdTe quantum dots analyzed by the chip-CE technique