Laboratory for Biochemistry and Glycobiology, Department of Biotechnology, Ghent University, 9000, Ghent, Belgium.
Laboratory of Biologically Active Molecules, Department of Biochemistry and Molecular Biology, Federal University of Ceara, 60455-760, Fortaleza, Brazil.
Glycoconj J. 2023 Feb;40(1):85-95. doi: 10.1007/s10719-022-10082-8. Epub 2022 Oct 26.
The Dalbergieae lectin group encompasses several lectins with significant differences in their carbohydrate specificities and biological properties. The current work reports on the purification and characterization of a GalNAc/Gal-specific lectin from Vataireopsis araroba (Aguiar) Ducke, designated as VaL. The lectin was purified from the seeds in a single step using guar gum affinity chromatography. The lectin migrated as a single band of about 35 kDa on SDS-PAGE and, in native conditions, occurs as a homodimer. The purified lectin is stable at temperatures up to 60 °C and in a pH range from 7 to 8 and requires divalent cations for its activity. Sugar-inhibition assays demonstrate the lectin specificity towards N-acetyl-D-galactosamine, D-galactose and related sugars. Furthermore, glycan array analyses show that VaL interacts preferentially with glycans containing terminal GalNAc/Galβ1-4GlcNAc. Biological activity assays were performed using three insect cell lines: CF1 midgut cells from the spruce budworm Choristoneura fumiferana, S2 embryo cells from the fruit fly Drosophila melanogaster, and GutAW midgut cells from the corn earworm Helicoverpa zea. In vitro assays indicated a biostatic effect for VaL on CF1 cells, but not on S2 and GutAW cells. The lectin presented a biostatic effect by reducing the cell growth and inducing cell agglutination, suggesting an interaction with glycans on the cell surface. VaL has been characterized as a galactoside-specific lectin of the Dalbergieae tribe, with sequence similarity to lectins from Vatairea and Arachis.
Dalbergieae 凝集素组包含几种凝集素，它们在碳水化合物特异性和生物学特性方面存在显著差异。本工作报道了从 Vataireopsis araroba（Aguiar）Ducke 中纯化和表征一种半乳糖/半乳糖特异性凝集素，命名为 VaL。该凝集素从种子中经瓜尔胶亲和层析一步纯化得到。SDS-PAGE 上该凝集素迁移为约 35 kDa 的单带，在天然条件下以同源二聚体形式存在。纯化的凝集素在高达 60°C 的温度和 pH 值为 7 至 8 的范围内稳定，并且需要二价阳离子才能发挥活性。糖抑制实验表明该凝集素对 N-乙酰-D-半乳糖胺、D-半乳糖和相关糖具有特异性。此外，糖芯片分析表明 VaL 优先与含有末端 GalNAc/Galβ1-4GlcNAc 的聚糖相互作用。使用三种昆虫细胞系：云杉卷叶蛾 Choristoneura fumiferana 的 CF1 中肠细胞、黑腹果蝇 Drosophila melanogaster 的 S2 胚胎细胞和玉米穗虫 Helicoverpa zea 的 GutAW 中肠细胞进行了生物活性测定。体外实验表明 VaL 对 CF1 细胞具有生物抑制作用，但对 S2 和 GutAW 细胞没有作用。该凝集素通过降低细胞生长和诱导细胞聚集呈现生物抑制作用，表明与细胞表面的聚糖相互作用。VaL 被表征为 Dalbergieae 部落的半乳糖苷特异性凝集素，与 Vatairea 和 Arachis 的凝集素有序列相似性。
