Isolation of a cytotoxic glycoprotein from the Scyphozoa Cyanea lamarckii by lectin-affinity chromatography and characterization of molecule interactions by surface plasmon resonance
Isolation of a cytotoxic glycoprotein from the Scyphozoa Cyanea lamarckii by lectin-affinity chromatography and characterization of molecule interactions by surface plasmon resonance
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DOI：
10.1016/j.jchromb.2008.06.040
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发表时间：
2008-08-01
影响因子：
3
通讯作者：
Prange, Andreas
中科院分区：
文献类型：
--
作者：
Helmholz, Heike;Naatz, Stefanie;Prange, Andreas
本发明描述了一种基于生物特异性凝集素亲和性的从远洋水母Cyanea lamarkii毒液中分离新型糖蛋白（ClGp 1）的方法，并且根据大小、分子相互作用和毒性对分离的糖蛋白进行化学和生物学表征。通过基质辅助激光解吸电离-飞行时间质谱（MALDI-TOF）测定，分离的蛋白质的分子量为25.7 kDa。酶促去糖基化后计算的碳水化合物含量为6.85 kDa。该糖蛋白具有细胞毒性，可从肠系膜和钓鱼触手的刺胞中分离。用表面等离子体共振（SPR）分析了该糖蛋白与凝集素伴刀豆球蛋白A（ConA）和麦胚凝集素（WGA）的结合行为，得到了对ConA的K（D）= 3.0 × 10 ~（-7）M和对WGA的K（D）= 2.1 × 10 ~（-6）M（pH5.0）和2.6 × 10 ~（-6）M（pH7.4）的亲和常数。(C)2008 Elsevier B. V.保留所有权利。
A biospecific lectin-affinity-based isolation process for a novel glycoprotein (ClGp1) from the venom of the pelagic jellyfish Cyanea lamarckii, is described and the isolated glycoprotein is chemically and biologically characterized according to size, molecular interaction and toxicity. The molecular mass of the isolated protein is 25.7 kDa as determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF). The carbohydrate content was calculated after enzymatic deglycosylation as 6.85 kDa. The glycoprotein is cytotoxic and could be isolated from cnidocysts of mesenteric and fishing tentacles. The binding behaviour of the glycoprotein to the lectins Concanavalin A (ConA) and Wheat Germ Agglutinin (WGA) was analyzed by surface plasmon resonance (SPR) and affinity constants in the range of K(D) = 3.0 x 10(-7) M for ConA and 2.1 x 10(-6) M (pH 5.0) and 2.6 x 10(-6) M (pH 7.4) for WGA were obtained. (C) 2008 Elsevier B.V. All rights reserved.