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四膜虫和梨形四膜虫中的镉硫蛋白。

Cadmium-thionein in Tetrahymena thermophila and Tetrahymena pyriformis.

机构信息

Dipartimento di Biologia, Università di Padova, Padova, Italia.

出版信息

Eur J Protistol. 1990 Oct 19;26(2):176-81. doi: 10.1016/S0932-4739(11)80112-4. Epub 2011 Nov 2.

Abstract

The treatment of Tetrahymena thermophila with cadmium causes a reduction in growth rate according to dose; almost all the metal is accumulated in the cytosol where the Zn content is also increased threefold. Bio-Gel and Water 160 (HPLC) column chromatography show that Cd and Zn are bound to a protein with an ultraviolet (UV) spectrum that appears to be similar to that of Cd-metallothioneins isolated by higher organisms, but its molecular weight is greater: about 28 000 D, comparable to that of metallothionein isolated from Tetrahymena pyriformis. Further purification of these proteins by ion exchange chromatography revealed the presence of two peaks, considered as two isoforms of the metallothioneins present in both T. thermophila and T. pyriformis (MT 1 and MT 2). Their amino acid analyses confirmed that they are different isometallothioneins, MT 1 and MT 2, with about 30% cysteine, and aspartic acid, glycine and lysine as major amino acids. From our analyses we may conclude that Tetrahymena pyriformis MTs are similar to those present in invertebrates and vertebrates, while Tetrahymena thermophila MTs are peculiar in that they have cyclic amino acid histidine in both MT 1 and MT 2; furthermore, aromatic amino acid phenylalanine is also present in MT 2.

摘要

用镉处理嗜热四膜虫会导致其生长速度按照剂量减少;几乎所有的金属都积累在细胞质中,锌的含量也增加了两倍。生物凝胶和水 160(HPLC)柱层析表明,Cd 和 Zn 与一种蛋白质结合,该蛋白质的紫外(UV)光谱似乎与从高等生物中分离出的 Cd-金属硫蛋白相似,但分子量更大:约 28000D,与从梨形四膜虫中分离出的金属硫蛋白相当。通过离子交换层析进一步纯化这些蛋白质,发现存在两个峰,被认为是存在于嗜热四膜虫和梨形四膜虫中的两种金属硫蛋白(MT1 和 MT2)的同工型。它们的氨基酸分析证实它们是不同的同工金属硫蛋白,MT1 和 MT2,约有 30%的半胱氨酸,天冬氨酸、甘氨酸和赖氨酸是主要的氨基酸。从我们的分析中可以得出结论,梨形四膜虫的金属硫蛋白与无脊椎动物和脊椎动物中的金属硫蛋白相似,而嗜热四膜虫的金属硫蛋白则很特别,因为它们在 MT1 和 MT2 中都有环状氨基酸组氨酸;此外,芳香族氨基酸苯丙氨酸也存在于 MT2 中。

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