[蓖麻毒素A链的溶酶体降解研究]
[Study on lysosomes degradation of ricin A chain].
作者信息
Chen Chun, Zhan Jin-biao, Shen Fen-ping, Shen Jian-gen
机构信息
Department of Biochemistry and Molecular Biology, College of Medicine, Zhejiang University, Hangzhou 310006, China.
出版信息
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2005 May;34(3):212-6. doi: 10.3785/j.issn.1008-9292.2005.03.004.
OBJECTIVE
To study lysosomes involvement in the degradation of ricin A chain.
METHODS
A lysosome-targeted singal KFERQ was added to the C terminus of rRTA by DNA recombinant technology. A pKK223.3 expression system in E. coli was used to produce recombinant ricine A chain (rRTA) and rRTA-KFERQ. Recombinant proteins were purified by affinity chromatography using Blue-Sepharose 6B. The cytotoxicity of recombinant proteins was measured by the MTT method.
RESULTS
Recombinant RTA-KFERQ was 49.87%, 54.18% and 88.68% less cytotoxic than RTA itself on the three cell lines HEPG2, Hela and A549, respectively.
CONCLUSION
Lysosomes can degrade, but not completely inactivate RTA in different cells, suggesting cells may have other degradation pathways for RTA.
目的
研究溶酶体在蓖麻毒素A链降解过程中的作用。
方法
通过DNA重组技术将溶酶体靶向信号KFERQ添加到rRTA的C末端。利用大肠杆菌中的pKK223.3表达系统生产重组蓖麻毒素A链（rRTA）和rRTA-KFERQ。重组蛋白通过使用Blue-Sepharose 6B的亲和色谱法进行纯化。采用MTT法测定重组蛋白的细胞毒性。
结果
重组RTA-KFERQ在HEPG2、Hela和A549这三种细胞系上的细胞毒性分别比RTA本身低49.87%、54.18%和88.68%。
结论
溶酶体可以降解，但不能完全使不同细胞中的RTA失活，这表明细胞可能存在RTA的其他降解途径。
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