Allen H J, Johnson E A
Carbohydr Res. 1977 Oct;58(2):253-65. doi: 10.1016/s0008-6215(00)84352-9.
L-Fucose-binding lectins from Ulex europeaus and Lotus tetragonolobus were isolated by affinity chromatography on columns of L-fucose-Sepharose 6B. L-Fucose was coupled to Sepharose 6B after divinyl sulfone-activation of the gel to give an affinity adsorbent capable of binding more than 1.2 mg of Ulex lextin/ml of gel, which could then be eluted with 0.1M or 0.05M L-fucose. Analysis of the isolated lectins by hemagglutination assay, by gel filtration, and polyacrylamide disc-electrophoresis revealed the presence of isolectins, or aggregated species, or both. The apparent mol. wt. of the major lectin fraction from Lotus was 35000 when determined on Sephadex G-200 or Ultrogel AcA 34. In contrast, the apparent mol. wt. of the major lectin fraction from Ulex was 68 000 when chromatographed on Sephadex G-200 and 45 000 when chromatographed on Ultrogel AcA 34. The yields of lectins were 4.5 mg/100 g of Ulex seeds and 394 mg/100 g of Lotus seeds.
通过在L-岩藻糖-琼脂糖6B柱上进行亲和层析,从欧洲荆豆和四角百脉根中分离出L-岩藻糖结合凝集素。在凝胶经二乙烯砜活化后,将L-岩藻糖偶联到琼脂糖6B上,得到一种亲和吸附剂,其每毫升凝胶能够结合超过1.2毫克的荆豆凝集素,然后可用0.1M或0.05M的L-岩藻糖洗脱。通过血凝试验、凝胶过滤和聚丙烯酰胺圆盘电泳对分离出的凝集素进行分析,结果显示存在同工凝集素、聚集物或两者皆有。在Sephadex G-200或Ultrogel AcA 34上测定时,来自百脉根的主要凝集素组分的表观分子量为35000。相比之下,在Sephadex G-200上进行层析时,来自荆豆的主要凝集素组分的表观分子量为68000,而在Ultrogel AcA 34上进行层析时为45000。凝集素的产量分别为每100克荆豆种子4.5毫克和每100克百脉根种子394毫克。