Implication of the differential roles of metallothionein 1 and 2 isoforms in the liver of rats as determined by polyacrylamide-coated capillary zone electrophoresis

Capillary electrophoresis of proteins 2003–2005

“…Linear polyacrylamide has been used to eliminate EOF in both fused-silica capillary [74][75][76] and channels in PDMS microchips [77]. Stability of the polyacrylamide coating was enhanced by cross-linking [78].…”

“…CZE was used to analyze protein isoforms including a 1 -antitrypsin (Fig. 4) [168], metallothionein isoforms in rat liver [76] and in Synecho- Figure 4. Four replicate CZE separations of six major a 1 -antitrypsin isoforms.…”

An Improved Diagnostic Method for Chronic Hepatic Disorder: Analyses of Metallothionein Isoforms and Trace Metals in the Liver of Patients with Hepatocellular Carcinoma as Determined by Capillary Zone Electrophoresis and Inductively Coupled Plasma-Mass Spectrometry

“…37,38) On the contrary, induction of MT-2 was reportedly greater than that of MT-1 in the liver of Zn-treated mice or tumor cells, 39) and Cu containing MT-2 induction was enhanced to a greater extent than that of MT-1 in the liver of LEC rats with the development of hepatitis. 24) In addition, commercially available MT-1 and MT-2 standards were observed to contain sub-isoforms. 40) Human hepatic MT isoforms in the tumor and juxta-tumor portion were distinctly separated into two isomers, MT-1 and MT-2, by capillary zone electrophoresis (Fig.…”

“…24) In order to suppress the protein adsorption and restrict the reversal of osmotic flow, the inner wall of the fused-silica capillary tube was coated with polyacrylamide by the method of Minami et al [28][29][30][31] The instrument conditions were as follows: the running buffer, 50 mM HEPES-Tris buffer (pH 7.4); samples, loaded onto the column for 7 s at a pressure of 50 mbar; the inlet of the capillary, anode; voltage, Ϫ25 kV; temperature, 26°C; detection, 214 and 254 nm. Samples were ultrafiltrated at molecular weight 50000 before running.…”

“…23) Although the biological functions and physiological role of MT are not yet known, based on the results of the analysis of hepatic MT-1 and MT-2 in LEC rats we recently proposed that MT-1 is related to the metabolism or detoxification of toxic metals such as cadmium, and, MT-2 is responsible for the homeostasis of essential metals such as Cu. 24) Since the pathophysiological feature of hepatic disorders in LEC rats has been reported to be very similar to that of human hepatic diseases involving HCC, 25,26) we attempted to determine MT-1 and MT-2 in the liver cytosol of humans by using a capillary zone electrophoresis (CZE). Previously, we had proposed the effectiveness of the multivariate discriminate analysis (MDA) when the three parameters, Cu, Zn, and MT were used.…”