[Separation and identification of metallothionein isoforms and sub-isoforms by liquid chromatography].
A method has been established for the separation and identification of metallothionein (MT) isoforms and sub-isoforms by liquid chromatography. A mixture of rabbit liver MT was separated on a DEAE A-25 weak anion-exchange chromatographic (AEC) column. Inductively coupled plasma-mass spectrometer (ICP-MS) was used for off-line detection of Zn in each fraction. The two main MT isoforms, MT-1 and MT-2, were isolated. The collected MT isoform fractions were then desalted by a homemade Sephadex G-25 size exclusion chromatographic (SEC) column. Afterwards, the sample was separated on a C18 reversed-phase column with UV detection at 210 nm. Different separation conditions were discussed and several sub-isoforms of MT were well separated at pH 2.0. The sub-isoforms were finally characterized by on-line HPLC-ESI-MS. The isoforms and sub-isoforms were well separated on a reversed-phase column under optimized chromatographic conditions and the detection results of ESI-MS were in agreement with the data found in literature. The method developed can be well used for the separation of metallothionein isoforms and sub-isoforms.
Lu
Suge
S
Department of Chemistry and the Key Laboratory of Analytical Science of the Ministry of Education, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China. lusuge@sina.com.cn
Shen
Jincan
J
Zhuang
Zhixia
Z
Wang
Xiaoru
X
D000818:Animals; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D008099:Liver / Q000737:chemistry*; D008668:Metallothionein / Q000737:chemistry* / Q000302:isolation & purification*; D020033:Protein Isoforms; D011817:Rabbits
D020033:Protein Isoforms; D008668:Metallothionein