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Purification and characterization of Moschatin, a novel type Ⅰ ribosome-inactivating protein from the mature seeds of pumpkin (Cucurbita moschata),and preparation of its immunotoxin against human melanoma cells

摘要A novel ribosome-inactivating protein designated Moschatin from the mature seeds of pumpkin (Cucurbita moschata) has been successively purified to homogeneity, using ammonium sulfate precipitation, CM-cellulose 52 column chromatography, Blue Sepharose CL-6B Affinity column chromatography and FPLC size-exclusion column chromatography. Moschatin is a type 1 RIP with a pI of 9.4 and molecular weight of~29 kD. It is a rRNA Nglycosidase and potently blocked the protein synthesis in the rabbit reticulocyte lysate with a IC50 of 0.26 nM. Using the anti-human melanoma McAb Ng76, a novel immunotoxin Moschatin-Ng76 was prepared successfully and it efficiently inhibited the growth of targeted melanoma cells M21 with a IC50 of 0.04 nM, 1500 times lower than that of free Moschatin. The results implied that Moschatin could be used as a new potential anticancer agent.

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作者单位 State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences,Beijing 100080, China [1] Department of Veterinary Pathobiology, University of Missouri-Columbia, MO 65211, USA [2]
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DOI 10.1038/sj.cr.7290182
发布时间 2004-04-09(万方平台首次上网日期,不代表论文的发表时间)
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细胞研究(英文版)

细胞研究(英文版)

2003年13卷5期

369-374页

SCIMEDLINEISTICCSCDBP

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