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Cochinin B, a novel ribosome-inactivating protein from the seeds of Momordica cochinchinensis.

机译:Cochinin B,一种来自于苦瓜种子的新型核糖体失活蛋白。

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摘要

Cochinin B, a novel ribosome-inactivating protein (RIP) with a molecular weight of 28 kDa, was purified from the seeds of Momordica cochinchinensis (Cucurbitaceae). The isolation procedure entailed ammonium sulfate precipitation, cation-exchange chromatography on SP Sepharose column and size-exclusion chromatography on Superdex 75 column with a fast protein liquid chromatography (FPLC) system. The first twenty N-terminal amino acid residues of Cochinin B showed homology to type I RIPs from other Momordica species. The purified Cochinin B displayed a strong inhibitory activity on protein synthesis in the cell-free rabbit reticulocyte lysate system with IC50 of 0.36 nM. Furthermore, it exhibited N-glycosidase activity and cytotoxicity against Vero cell line with IC50 higher than 1540 nM. Interestingly, Cochinin B manifested strong anti-tumor activities on human cervical epithelial carcinoma (HeLa), human embryonic kidney (HEK293) and human small cell lung cancer (NCI-H187) cell lines with IC50 of 16.9, 114 and 574 nM, respectively.
机译:Cochinin B是一种分子量为28 kDa的新型核糖体失活蛋白(RIP),是从苦瓜木(葫芦科)的种子中纯化得到的。分离过程包括硫酸铵沉淀,SP Sepharose柱上的阳离子交换色谱和Superdex 75柱上的尺寸排阻色谱,以及快速蛋白液相色谱(FPLC)系统。 Cochinin B的前20个N末端氨基酸残基与来自其他苦瓜属的I型RIP具有同源性。纯化的Cochinin B在无细胞兔网织红细胞裂解液系统中显示出对蛋白质合成的强大抑制活性,IC50为0.36 nM。此外,它显示出对Vero细胞系的N-糖苷酶活性和细胞毒性,IC50高于1540 nM。有趣的是,Cochinin B对人宫颈上皮癌(HeLa),人胚胎肾脏(HEK293)和人小细胞肺癌(NCI-H187)细胞系表现出强大的抗肿瘤活性,IC50分别为16.9、114和574 nM。

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