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Purification and characterization of a lectin with antiproliferative activity toward cancer cells from the dried fruit bodies of Lactarius flavidulus

机译:一种从黄精乳干果体内对癌细胞具有抗增殖活性的凝集素的纯化和表征

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摘要

The isolation of a dimeric 29.8-kDa lectin (LFL) from dried Lactarius flavidulus fruit bodies is reported herein. The chromatographic procedure utilized comprised anion-exchange chromatography on DEAE-cellulose, cation-exchange chromatography on CM-cellulose, anion-exchange chromatography on SP-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of LFL was inhibited by a variety of simple sugars, such as lactose, p-nitrophenyl α-d-glucopyranoside, p-nitrophenyl β-d-glucopyranoside and inositol, and by the polysaccharide inulin. The activity of LFL was stable up to 40 °C. There was a precipitous drop in activity when the temperature was elevated to 50 °C. Hemagglutinating activity was retained when LFL was exposed to 6.25-12.5 mM HCl and NaOH. The activity was potently inhibited by Fe~(2+) and Fe ~(3+) ions, and slightly inhibited by Al~(3+) and Mn ~(2+) ions. LFL suppressed the proliferation of hepatoma (HepG2) and leukemic (L1210) cells with an IC_(50) of 8.90 and 6.81 μM, respectively. It inhibited the activity of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) with an IC_(50) of 5.68 μM. However, LFL did not exhibit antifungal activity.
机译:本文报道了从干燥的黄精乳杆菌子实体中分离出二聚体29.8-kDa凝集素(LFL)。使用的色谱方法包括:DEAE-纤维素上的阴离子交换色谱,CM-纤维素上的阳离子交换色谱,SP-Sepharose上的阴离子交换色谱以及Superdex 75上的快速蛋白液相色谱进行凝胶过滤。LFL的血凝活性为被各种简单的糖,例如乳糖,对硝基苯基α-d-吡喃葡萄糖苷,对硝基苯基β-d-吡喃葡萄糖苷和肌醇抑制,并被多糖菊粉抑制。 LFL的活性在最高40°C时稳定。当温度升高到50℃时,活性急剧下降。当LFL暴露于6.25-12.5 mM HCl和NaOH中时,血凝活性得以保留。 Fe〜(2+)和Fe〜(3+)离子对活性的抑制作用强,而Al〜(3+)和Mn〜(2+)离子对活性的抑制作用很小。 LFL抑制肝癌细胞(HepG2)和白血病(L1210)细胞的增殖,IC_(50)分别为8.90和6.81μM。它以5.68μM的IC_(50)抑制1型人类免疫缺陷病毒(HIV-1)逆转录酶(RT)的活性。但是,LFL没有表现出抗真菌活性。

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