Rapid identification of metallothionein isoforms in liver cytosol fraction by capillary zone electrophoresis using EDTA.
Identification of metallothionein (MT) isoforms on capillary zone electrophoresis (CZE) analysis was studied using a linear polyacrylamide-coated capillary at pH 7.4 and EDTA. The CZE system was able to separate standard (purified and commercially available) MT specimens into their isoforms within 10 min. The peaks of MT-1 and MT-2 isoforms disappeared on addition of EDTA to the specimen, and the disappearance was shown to be time-dependent and dose-dependent, although the reason why the peaks decreased is still unclear. A heat-treated cytosol fraction prepared from Zn-injected mouse liver showed many major and minor peaks on CZE analysis. Two major peaks were identified to be MT-1 and MT-2, respectively, by co-injection with the purified MT isoforms. When EDTA was added to the cytosol fraction, the two major peaks, MT-1 and MT-2, and three other minor peaks disappeared time-dependently. Therefore, each MT isoform in the cytosol fraction can be identified by the addition of EDTA, also the peaks are identified by the corresponding migration times of purified MTs. Unknown substances like MT sub-isoforms may also be detected, although this question warrants clarification. From these results, it was concluded that the addition of EDTA is useful for identification of MT isoforms in cytosol fractions on CZE analysis.
Kubo
K
K
Faculty of Pharmaceutical Sciences, Kinki University, Osaka, Japan.
Sakita
Y
Y
Otaki
N
N
Kimura
M
M
Minami
T
T
D000818:Animals; D003600:Cytosol / Q000737:chemistry; D004492:Edetic Acid; D019075:Electrophoresis, Capillary / Q000379:methods*; D008099:Liver / Q000737:chemistry*; D008668:Metallothionein / Q000032:analysis*; D051379:Mice; D020033:Protein Isoforms / Q000032:analysis*
D020033:Protein Isoforms; D008668:Metallothionein; D004492:Edetic Acid