Departamento de Bioquímica-CCB, Universidade Federal de Pernambuco, Cidade Universitária, 50670-420 Recife, Pernambuco, Brazil.
Plant Sci. 2012 Feb;183:20-6. doi: 10.1016/j.plantsci.2011.10.018. Epub 2011 Nov 4.
Hemagglutinating activity has been associated to presence of lectin, carbohydrate-binding proteins. In this work Crataeva tapia bark lectin (CrataBL) was purified in milligram quantities (28 mg per g of bark) by ion exchange chromatography. The lectin was thermo-stable, ion-independent and N-terminal sequence analysis demonstrated similarity with miraculin and miraculin-like proteins (plant defensive proteins). Glycosylated nature of CrataBL was revealed using glycoprotein staining (periodic acid-Schiff's reagent), positive for polypeptides of apparent molecular masses 21 and 40 kDa on SDS-PAGE. Gel diffusion assay showed that glucose/mannose isolectins from Cratylia mollis recognized CrataBL glycan moiety. CrataBL hemagglutinating activity was inhibited by glycoproteins and CrataBL immobilized on cyanogen bromide-activated sepharose 4B (1 mL) bound 0.54 mg of glycoprotein (casein, fetuin and ovalbumin) per cycle. CrataBL was an insecticide agent against Nasutitermes corniger workers (termite that attack woods) with LC₅₀ of 0.475 mg mL⁻¹ for 6 days.
血凝活性与凝集素（碳水化合物结合蛋白）的存在有关。在这项工作中，通过离子交换层析从 Crataeva tapia 树皮中以毫克级（每克树皮 28 毫克）的量纯化了 Crataeva tapia 树皮凝集素（CrataBL）。该凝集素热稳定，离子独立，N 末端序列分析表明与奇迹素和奇迹素样蛋白（植物防御蛋白）具有相似性。使用糖蛋白染色（过碘酸-Schiff 试剂）揭示了 CrataBL 的糖基化性质，在 SDS-PAGE 上显示出 21 和 40 kDa 左右的多肽为阳性。凝胶扩散测定表明，来自 Cratylia mollis 的葡萄糖/甘露糖同凝集素识别 CrataBL 的糖基部分。CrataBL 的血凝活性被糖蛋白抑制，并且固定在氰基溴化活化的 sepharose 4B 上的 CrataBL（1 mL）每轮结合 0.54 mg 糖蛋白（酪蛋白、胎球蛋白和卵清蛋白）。CrataBL 是一种针对 Nasutitermes corniger 工蚁（攻击木材的白蚁）的杀虫剂，6 天的 LC₅₀为 0.475 mg mL⁻¹。
