[具有生物活性的重组蓖麻毒素B链的制备]

[Production of biologically active recombinant ricin B-chain].

作者信息

Tonevitskiĭ A G, Toptygin A Iu, Agapov I I, Rakhmanova V A, Shamshiev A T, Alekseev Iu O, Pfueller U, Frankel A

机构信息

Institute for Phytochemistry, University of Witten/Herdecke, Witten, Germany.

出版信息

Mol Biol (Mosk). 1995 Mar-Apr;29(2):398-406.

PMID:7783743

Abstract

Escherichia coli cells transformed with plasmids containing ricin B-chain coding sequences are shown to express this heterologous protein in inclusion bodies. After denaturation and renaturation of the product in the presence of glutathione and lactose, the recombinant ricin B-chain is soluble, biologically active and stable. Cytotoxicity of heterodimer containing this protein and ricin A-chain is found to be only ten times lower, than that of native ricin. Recombinant B-chain alone was nontoxic to cells (ID50 > 10(-6) M). Our data suggest that ricin B-chain oligosaccharides are essential for stability preserving protein from proteolytic degradation in cells.

摘要

用含有蓖麻毒素B链编码序列的质粒转化的大肠杆菌细胞在包涵体中表达这种异源蛋白。在谷胱甘肽和乳糖存在下对产物进行变性和复性后，重组蓖麻毒素B链是可溶的、具有生物活性且稳定的。发现含有这种蛋白和蓖麻毒素A链的异源二聚体的细胞毒性仅比天然蓖麻毒素低10倍。单独的重组B链对细胞无毒（半数抑制浓度>10(-6) M）。我们的数据表明，蓖麻毒素B链寡糖对于保护蛋白质免于细胞内蛋白水解降解的稳定性至关重要。