高等学校化学学报

高等学校化学学报 ›› 1993, Vol. 14 ›› Issue (4): 454.

• 研究论文 • 上一篇    下一篇

大鼠肝脏金属硫蛋白的分离纯化和金属硫蛋白亚型-Ⅱα结构域片段的制备及其核磁共振鉴定

黄仲贤, 顾伟强, 胡红雨, 王韵华   

  1. 复旦大学化学系, 上海 200433
  • 收稿日期:1992-07-28 修回日期:1992-10-11 出版日期:1993-04-24 发布日期:1993-04-24
  • 通讯作者: 黄仲贤
  • 基金资助:

    国家自然科学基金

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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摘要： Wistar雄性大鼠经腹腔注射CdCl2诱导其肝脏合成金属硫蛋白。取鼠肝,先经匀浆、热变性、乙醇-氯仿萃取、丙酮沉淀等步骤,后由SephadexG-75、DEAE-52柱层析,得金属硫蛋白的两亚型Ⅰ和Ⅱ。经原子吸收光谱测定每蛋白分子含有5个镉和2个锌。用枯草杆菌酶割法制备并分离了金属硫蛋白亚型Ⅱ的α结构域片段。还用NMR研究了金属硫蛋白的两个亚型及所得的α结构域片段。

关键词: Cd5Zn2-金属硫蛋白, 大鼠肝脏金属硫蛋白, 金属硫蛋白的&alpha, 结构域片段, 蛋白质NMR

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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