Lectins from tropical sponges -: Purification and characterization of lectins from genus Aplysina
Lectins from tropical sponges -: Purification and characterization of lectins from genus Aplysina
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DOI：
10.1074/jbc.m001366200
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发表时间：
2000-09-22
影响因子：
4.8
通讯作者：
Fresno, M
中科院分区：
文献类型：
--
作者：
Miarons, PB;Fresno, M
只有少数动物门已被筛选的凝集素的存在和分布，可能是最深入的研究组是软体动物。在这项调查中，22种从12个家庭的热带海绵收集在洛斯罗格国家公园（委内瑞拉）进行了筛选凝集素的存在。9种生理盐水浸提液对链霉蛋白酶处理的仓鼠红细胞表现出较强的血凝活性;其中5种与兔红细胞反应，4种与胰蛋白酶处理的牛红细胞反应，5种与人红细胞反应，无论血型类型如何。从Aplysina属（archeri，lawnosa和cauliformis）研究的三个物种的提取物具有高度反应性，并对测试的红细胞组具有泛凝集性。A. archeri和A.通过硫酸铵分级分离、对氨基苄基-β-1-硫代吡喃半乳糖苷-琼脂糖亲和层析和凝胶过滤层析将草坪草纯化至均一。这两种凝集素均表现出63 kDa的天然分子量，并通过还原条件下的SDS-聚丙烯酰胺凝胶电泳具有16 kDa的表观分子量，从而表明它们以同源四聚体形式存在。纯化的凝集素每分子含有3-4摩尔的二价阳离子，这是它们的生物活性所必需的。用半抗原抑制血凝试验确定纯化的A. archeri凝集素。结果表明，凝集素对非还原性β-连接的D-Gal残基的偏好是红细胞结合甲基-β-D-Gal和硫代二半乳糖苷（Gal β 1-4-硫代半乳糖吡喃糖苷）的最佳抑制剂。几种聚糖在固定化凝集素亲和色谱上的行为证实并扩展了通过半抗原抑制获得的特异性数据。
Only a few animal phyla have been screened for the presence and distribution of lectins, Probably the most intensively studied group is the mollusk. In this investigation, 22 species from 12 families of tropical sponges collected in Los Rogues National Park (Venezuela) were screened for the presence of lectins. Nine saline extracts exhibited strong hemagglutinating activity against pronase-treated hamster red blood cells; five of these reacted against rabbit red blood cells, four with trypsin-treated bovine red blood cells, and five with human red blood cells regardless of the blood group type. Extracts from the three species studied from genus Aplysina (archeri, lawnosa, and cauliformis) were highly reactive and panagglutinating against the panel of red blood cells tested. The lectins from A. archeri and A. lawnosa were purified to homogeneity by ammonium sulfate fractionation, affinity chromatography on p-aminobenzyl-beta-1-thiogalactopyranoside-agarose, and gel filtration chromatography. Both lectins exhibited a native molecular mass of 63 kDa and by SDS-polyacrylamide gel electrophoresis under reducing conditions have an apparent molecular mass of 16 kDa, thus suggesting they occur as homotetramers. The purified lectins contain 3-4 mol of divalent cation per molecule, which are essential for their biological activity. Hapten inhibition of hemagglutination was carried out to define the sugar binding specificity of the purified A. archeri lectin. The results indicate a preference of the lectin for nonreducing beta-linked D-Gal residues being the best inhibitors of red blood cells binding methyl-beta-D-Gal and thiodigalactoside (Gal beta 1-4-thiogalactopyranoside). The behavior of several glycans on immobilized lectin affinity chromatography confirmed and extended the specificity data obtained by hapten inhibition.