A lactose specific lectin from the sponge Cinachyrella apion: Purification, characterization, N-terminal sequences alignment and agglutinating activity on Leishmania promastigotes
A lactose specific lectin from the sponge Cinachyrella apion: Purification, characterization, N-terminal sequences alignment and agglutinating activity on Leishmania promastigotes
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DOI：
10.1016/j.cbpb.2009.10.016
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发表时间：
2010-03-01
影响因子：
2.2
通讯作者：
Santos, Elizeu A.
中科院分区：
文献类型：
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作者：
Medeiros, Danielle S.;Medeiros, Thales L.;Santos, Elizeu A.
海绵Cinachyrella apion的粗提物显示出与多克隆抗体IgG抗CvL（Cliona varians lectin）的交叉反应性，并且对所有ABO组的人红细胞也具有很强的血细胞凝集活性。因此，在Superose 6 10/300柱上进行丙酮分级分离、IgG抗去糖基化CvL琼脂糖亲和层析和快速蛋白质液相层析（FPLC-AKTA Purifier）凝胶过滤以纯化新的凝集素。C.蜜蜂凝集素（CaL）凝集所有类型的人红细胞，优先木瓜蛋白酶化的A型红细胞。血细胞凝集活性不依赖于Ca 2+、Mg 2+和Mn 2+离子，并且它被二糖乳糖强烈抑制，最低浓度为6.25 mM。通过在Superose 12 10/300柱上的FPLC-凝胶过滤和SDS凝胶电泳测定的CaL分子量约为124 kDa，由8个15.5 kDa的亚基组成，通过疏水相互作用组装。该凝集素在0至60 ℃之间是热稳定的，并且pH稳定。对CaL的N-末端氨基酸序列进行了测定，并对氨基酸序列进行了Blast搜索，结果显示该蛋白仅与一个silicatein具有相似性。恰加斯利什曼原虫前鞭毛体被Cal凝集，这种活性被乳糖消除，表明乳糖受体可以在该寄生虫阶段呈现。这些发现表明，潜在的生物技术应用卡尔。作为诊断病原性原生动物。(C)2009 Elsevier Inc. All rights reserved.
Crude extract from the sponge Cinachyrella apion showed cross-reactivity with the polyclonal antibody IgG anti-CvL (Cliona varians lectin) and also a strong haemagglutinating activity towards human erythrocytes of all ABO groups. Thus, it was submitted to acetone fractionation, IgG anti-deglycosylated CvL Sepharose affinity chromatography, and Fast Protein Liquid Chromatography (FPLC-AKTA Purifier) gel filtration on a Superose 6 10/300 column to purify a novel lectin. C. apion lectin (CaL) agglutinated all types of human erythrocytes with preference for papainized type A erythrocytes. The haemagglutinating activity is independent of Ca2+, Mg2+ and Mn2+ ions, and it was strongly inhibited by the disaccharide lactose, up to a minimum concentration of 6.25 mM. CaL molecular mass, determined by FPLC-gel filtration on a Superose 12 10/300 column and SDS gel electrophoresis, was approximately 124 kDa, consisting of eight subunits of 15.5 kDa, assembled by hydrophobic interactions. The lectin was heat-stable between 0 and 60 degrees C and pH-stable. The N-terminal amino acid sequence of CaL was also determined and a blast search on amino acid sequences revealed that the protein showed similarity only with a silicatein. Leishmania chagasi promastigotes were agglutinated by Cal. and this activity was abolished by lactose, indicating that lactose receptors could be presented in this parasite stage. These findings are indicative of the potential biotechnological application of Cal. as diagnostic of pathogenic protozoa. (C) 2009 Elsevier Inc. All rights reserved.