Isolation and characterization of a novel lectin from the wild mushroom Xerocomus spadiceus
Isolation and characterization of a novel lectin from the wild mushroom Xerocomus spadiceus
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DOI：
10.1016/j.peptides.2003.11.013
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发表时间：
2004-01-01
期刊：
影响因子：
3
通讯作者：
Ng, TB
中科院分区：
文献类型：
--
作者：
Liu, QH;Wang, HX;Ng, TB
使用涉及DEAE-纤维素上的离子交换色谱法、Affi-gel蓝色凝胶上的亲和色谱法、CM-纤维素上的离子交换色谱法以及Superdex上的快速蛋白质液相色谱法的凝胶过滤的程序，从蘑菇Xerocomus spadiceus的子实体提取物中分离凝集素。 75. 凝集素未吸附在 DEAE-纤维素和 Affi-gel 蓝色凝胶上，而吸附在 CM-Sepharose 上。凝集素能够在小鼠脾细胞中引发大约四倍的有丝分裂反应刺激。血凝活性在60℃以下保持稳定，在70℃时减半，在75℃时降低至25％，在80℃时降低至不可检测的水平。在盐浓度高达 10 mM 的各种二价（Ca2+、Mg2+、Zn2+ 和 Mn2+）氯化物存在下，活性保持不变（ZnCl2 除外）。 FeCl3 浓度高达 10 mM 时不会影响凝集素的血凝活性。凝集素的血凝活性在 5 mM AlCl3 或 10 mM ZnCl2 存在下加倍，在 10 mM AlCl3 存在下四倍。当 HCl 和 NaOH 存在时，活性降低。在所测试的大量碳水化合物中，只有菊粉能够抑制凝集素的血凝活性。 (C) 2004 Elsevier Inc. 保留所有权利。
A lectin was isolated from extracts of fruiting bodies of the mushroom Xerocomus spadiceus using a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose, and gel filtration by fast protein liquid chromatography on Superdex. 75. The lectin was unadsorbed on DEAE-cellulose and Affi-gel blue gel and adsorbed on CM-Sepharose. The lectin was capable of eliciting an approximately four-fold stimulation of mitogenic response in murine splenocytes. The hemagglutinating activity was stable up to 60degreesC, halved at 70degreesC, reduced to 25% at 75degreesC and dwindled to an undetectable level at 80degreesC. The activity remained unaltered in the presence of various divalent (Ca2+, Mg2+, Zn2+ and Mn2+) chlorides up to a salt concentration of 10 mM except in the case of ZnCl2. FeCl3 up to a concentration of 10 mM did not affect the hemagglutinating activity of the lectin. The hemagglutinating activity of the lectin was doubled in the presence of 5 mM AlCl3 or 10 mM ZnCl2 and quadrupled in the presence of 10 mM AlCl3. The activity was reduced in the presence of HCl and NaOH. Among the large number of carbohydrates tested, only inulin was able to inhibit the hemagglutinating activity of the lectin. (C) 2004 Elsevier Inc. All rights reserved.