Helmholz Heike, Naatz Stefanie, Lassen Stephan, Prange Andreas
GKSS Research Centre Geesthacht, Institute for Coastal Research, Department for Marine Bioanalytical Chemistry, Geesthacht, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Aug 1;871(1):60-6. doi: 10.1016/j.jchromb.2008.06.040. Epub 2008 Jul 1.
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.
本文描述了一种基于生物特异性凝集素亲和的方法，用于从远洋水母拉氏霞水母（Cyanea lamarckii）的毒液中分离一种新型糖蛋白（ClGp1），并根据大小、分子相互作用和毒性对分离出的糖蛋白进行了化学和生物学表征。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）测定，分离出的蛋白质分子量为25.7 kDa。酶促去糖基化后计算出碳水化合物含量为6.85 kDa。该糖蛋白具有细胞毒性，可从肠系膜和捕食触手的刺丝囊中分离得到。通过表面等离子体共振（SPR）分析了该糖蛋白与凝集素刀豆球蛋白A（ConA）和麦胚凝集素（WGA）的结合行为，获得的亲和常数范围为：ConA的KD = 3.0 x 10(-7) M，WGA在pH 5.0时为2.1 x 10(-6) M，在pH 7.4时为2.6 x 10(-6) M。
