Structural changes in metallothionein isoforms revealed by capillary electrophoresis and Brdicka reaction
Jazyk angličtina Země Německo Médium print
Typ dokumentu časopisecké články
PubMed
22222973
DOI
10.1002/elps.201100312
Knihovny.cz E-zdroje
- MeSH
- biochemické jevy MeSH
- chemická precipitace MeSH
- chemické modely * MeSH
- denaturace proteinů MeSH
- elektroforéza kapilární metody MeSH
- králíci MeSH
- lineární modely MeSH
- metalothionein chemie metabolismus MeSH
- močovina chemie MeSH
- molekulární sekvence - údaje MeSH
- nádorové biomarkery analýza chemie MeSH
- protein - isoformy MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- senzitivita a specificita MeSH
- spektrofotometrie ultrafialová MeSH
- vysoká teplota MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- metalothionein MeSH
- močovina MeSH
- nádorové biomarkery MeSH
- protein - isoformy 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.
Citace poskytuje Crossref.org
Fluorescence-tagged metallothionein with CdTe quantum dots analyzed by the chip-CE technique