Structural changes in metallothionein isoforms revealed by capillary electrophoresis and Brdicka reaction

Abstract

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.

Improved Electrochemical Detection of Zinc Ions Using Electrode Modified with Electrochemically Reduced Graphene Oxide

“…As was shown recently, the enhanced zinc intake by drinking water in the case of mice caused zinc deficiency in the hippocampus, associated with memory deficit and decreased expression levels of learning and memory related receptors [5]. Zinc has these important roles and effects mainly as a co-factor of numerous proteins, therefore it is not surprising that metallomics and proteomics of zinc-containing proteins are emerging fields of science [6,7,8]. From these, metallothioneins are highlighted as maintainers and transporters of these proteins and their importance in zinc metabolism belongs to the interest of numerous researchers [9,10,11,12,13,14,15,16,17,18].…”

Bioconjugation of peptides using advanced nanomaterials to examine their interactions in 3D printed flow‐through device

“…MTs are currently considered to be highly important in cellular protection against oxidative stress, cytotoxic agents, bacterial infections, radiation, and/or DNA damage . The tertiary structure of MTs is based on the presence of two domains (α and β) that form cysteine clusters to bind metal ions and are responsible for MTs biological activity . Although several studies have demonstrated upregulation of MTs in organisms exposed to viruses, such as hepatitis C or influenza A , no evidences regarding the direct interactions between MTs and viral surface antigens are currently available.…”

Metallothionein gene expression in renal cell carcinoma

“…[13] New biological roles for these proteins have been identified, including those needed in the carcinogenic process. However, their use as a predictive marker remains controversial.…”