CJCU

Chem. J. Chinese Universities ›› 1993, Vol. 14 ›› Issue (4): 454.

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Separation and Purification of Rat Liver Metallothionein, Preparation of α-Domain Fragments of MT-Ⅱ and Their Characterization by NMR

HUANG Zhong-Xian, GU Wei-Qiang, HU Hong-Yu, WANG Yun-Hua   

  1. Department of Chemistry, Fudan University, Shanghai, 200433
  • Received:1992-07-28 Revised:1992-10-11 Online:1993-04-24 Published:1993-04-24

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Abstract: The rat liver metallothionein was induced by administering 7 subcutaneous injection of CdCl2 to the wistar rat.The purification protocal involved homogenization of rat liver, heating, extraction by ethanol-chloroform, acetone precipitation, and finally, chromatography on Sephadex G-75 and DEAE-52 columns.Two isoforms of metallothionein, Cd5Zn2-MT-Ⅰ and-Ⅱ, with A250/ A250> 20 were obtained.MT-Ⅱα domain fragment containing four cadmium ions was also isolated by digestion with subtilysin.1H NMRspectra of MT-Ⅰ,-Ⅱ were in agreement with those reported in literatures.The spectrum of MT-Ⅰ α domain shows that there is no existence of β fragment in this preparation.

Key words: Cd5Zn2-metallothionein, Rat liver metallothionein, α-domain fragment of metallothionein, Protein NMR

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