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化学学报
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化学学报 ›› 1998, Vol. 56 ›› Issue (11): 1135-1144. 上一篇    

研究论文

苦瓜子蛋白的分离纯化及其性质研究

叶国杰;钱瑞卿;卢保元;顾梓伟;金善炜;汪猷   

  1. 中国科学院上海有机化学研究所.上海(200032)
  • 发布日期:1998-11-15

Isolation and characterization of α, β-momorcharins

Ye Guojie;Qian Ruiqing;Lu Baoyuan;Gu Ziwei;Jin Shanwei;Wang You   

  1. Shanghai Inst Organ Chem., CAS.Shanghai(200032)
  • Published:1998-11-15

PDF

453

被引次数

30

从苦瓜种子的粗提物苦瓜子蛋白丙酮分级沉淀干粉中,用CM-SephadexC-50和SephadexG-75分离得到了两个核糖体失活蛋白,α-苦瓜子蛋白(α-momorcharin,α-MMC)和β-苦瓜子蛋白(β-momorcharin,β-MMC),其等电点分别为9.10(α-MMC),9.32(β-MMC)。用ESI-MS和MALDI-TOF-MS测定它们的分子量,分别为28625(α-MMC),29076(β-MMC)(+Na,29099);28795(α-MMC),29074.7(β-MMC)。它们都是糖蛋白。其生物活性的分析测定表明,都属于RNAN-糖苷酶。本文重点对β-苦瓜子蛋白的分离纯化及其性质进行详细报道,并对其N-端部分的氨基酸顺序进行了测定。

关键词: 苦瓜子蛋白, 分离, 纯化, 核糖体失活蛋白, 糖苷酶, 糖蛋白, 生物活性, 氨基酸顺序分析, 苦瓜, 葫芦科

Two ribosome-inactivating proteins (RIP's), α- and β- momorcharin (α- and β-MMC) have been isolated from the seeds of bitter melon (Momordica charantia), a traditional Chinese folk medicine. This was done by acetone fractionaction and chromatographies on CM-Sephadex C- 50 and Sephadex G-75. The molecular weights of the purified proteins as determined by ESI-MS and MALDI-TOF-MS were 28625 (α-MMC), 29076(β -MMC) and 28795 (α-MMC), 29074(β-MMC), respectively. Both of them were found to be glycoproteins and have pI values of 9.10 (α-MMC) and 9.32 (β-MMC). Biological activity assay revealed both of them were RNAN-glycosidase and depurinate 28S rRNA in the ribosomes in a manner identical to that of trichosanthin. Here the isolation, physicochemical properties and N-terminal amino acid sequence of β- MMC are described.

Key words: SEPARATION, PURIFICATION, GLYCOSIDASE, GLYCOPROTEIN, BIOLOGICAL ACTIVITY, AMINO ACID SEQUENCE ANALYSIS, MOMORDICA CHARANTIA, CUCURBITACEAE

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