Hosono M, Kawauchi H, Nitta K, Takayanagi Y, Shiokawa H, Mineki R, Murayama K
Cancer Research Institute, Tohoku College of Pharmaceutical Sciences, Sendai, Japan.
Biol Pharm Bull. 1993 Mar;16(3):239-43. doi: 10.1248/bpb.16.239.
Three rhamnose-binding lectins were purified from the roe of Osmerus eperlanus mordax (olive rainbow smelt) by affinity chromatography and ion-exchange chromatography. The apparent molecular weights of Osmerus eperlanus mordax lectin (OML) -1, -2 and -3 were 25000, 32000 and 26000, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. On native PAGE, these three lectins showed different migration patterns (Rm value; 0.37, 0.53 and 0.66, respectively). OMLs agglutinated rabbit and human type B erythrocytes and sarcoma 180 cells, but not human type A and O erythrocytes and AH109A cells. The most effective monosaccharide inhibitor was L-rhamnose. L-Mannose and D-galactose were also good inhibitors. Furthermore, OML-induced hemagglutination was inhibited more strongly by melibiose or raffinose rather than lactose or lactulose. Therefore, OMLs are L-rhamnose/alpha-D-galactosyl type lectins. OMLs did not require a detergent, when extracted from crude material, and Ca2+, Mg2+, EDTA and dithiothreitol were not necessary for the OML-induced hemagglutination activities. The OMLs had similar N-terminal amino acid sequences.
通过亲和色谱和离子交换色谱法从欧洲胡瓜鱼（橄榄色虹鳟）的鱼卵中纯化出三种鼠李糖结合凝集素。在还原条件下进行的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）中，欧洲胡瓜鱼凝集素（OML）-1、-2和-3的表观分子量分别为25000、32000和26000。在非变性PAGE上，这三种凝集素表现出不同的迁移模式（相对迁移率值分别为0.37、0.53和0.66）。OML能凝集兔和人的B型红细胞以及肉瘤180细胞，但不能凝集人的A型和O型红细胞以及AH109A细胞。最有效的单糖抑制剂是L - 鼠李糖。L - 甘露糖和D - 半乳糖也是良好的抑制剂。此外，蜜二糖或棉子糖对OML诱导的血凝作用的抑制比乳糖或乳果糖更强。因此，OML是L - 鼠李糖/α - D - 半乳糖基型凝集素。从粗原料中提取OML时不需要去污剂，并且Ca2 +、Mg2 +、EDTA和二硫苏糖醇对OML诱导的血凝活性不是必需的。这几种OML具有相似的N端氨基酸序列。
