Gupta D, Bhattacharyya L, Fant J, Macaluso F, Sabesan S, Brewer C F
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461.
Biochemistry. 1994 Jun 21;33(24):7495-504. doi: 10.1021/bi00190a001.
The tetrameric lectin from Glycine max (soybean) (SBA) has been shown to cross-link and precipitate with N-linked multiantennary complex type oligosaccharides containing nonreducing terminal Gal residues (Bhattacharyya, L., Haraldsson, M., & Brewer, C. F. (1988) Biochemistry 27, 1034-1041). In the present study, negative stain electron micrographs of the precipitates of SBA with a series of naturally occurring and synthetic multiantennary carbohydrates with terminal Gal or GalNAc residues show the presence of highly ordered cross-linked lattices for many of the complexes. The precipitates of SBA with a "bisected" and "nonbisected" N-linked biantennary complex type oligosaccharide containing Gal residues at the nonreducing termini show similar two-dimensional patterns. However, the pattern observed for the precipitates of a tetraantennary complex type oligosaccharide with SBA is distinct from those of the two biantennary carbohydrates. Furthermore, the precipitates formed between the lectin and a synthetic O-linked biantennary ("cluster") glycoside with terminal GalNAc residues show a pattern that is different from those above. Four biantennary pentasaccharide analogs of the blood group I antigen containing beta-LacNAc moieties at the 2,3-, 2,4-, 2,6-, and 3,6-positions of the core Gal also showed ordered patterns in their precipitates with SBA. X-ray crystallographic data and mixed quantitative precipitation profiles of binary mixtures of the four analogs demonstrate that each analog possesses a unique cross-linked lattice with the protein. A common structural feature of the naturally occurring and synthetic carbohydrates that show highly organized cross-linked lattices with SBA is the presence of a pseudo-2-fold axis of symmetry in each oligosaccharide relating the terminal binding epitopes on each arm. This suggests that the symmetry features of certain naturally occurring branch chain oligosaccharides facilitate formation of highly ordered, homogeneous cross-linked complexes with specific lectins.
来自大豆(Glycine max)的四聚体凝集素(SBA)已被证明能与含有非还原末端Gal残基的N-连接多天线复合型寡糖发生交联并沉淀(Bhattacharyya, L., Haraldsson, M., & Brewer, C. F. (1988) Biochemistry 27, 1034 - 1041)。在本研究中,SBA与一系列含有末端Gal或GalNAc残基的天然存在和合成的多天线碳水化合物沉淀的负染色电子显微照片显示,许多复合物存在高度有序的交联晶格。SBA与在非还原末端含有Gal残基的“平分型”和“非平分型”N-连接双天线复合型寡糖的沉淀显示出相似的二维模式。然而,四天线复合型寡糖与SBA沉淀所观察到的模式与两种双天线碳水化合物的不同。此外,凝集素与含有末端GalNAc残基的合成O-连接双天线(“簇”)糖苷之间形成的沉淀显示出与上述不同的模式。四种血型I抗原的双天线五糖类似物,在核心Gal的2,3-、2,4-、2,6-和3,6-位含有β-LacNAc部分,其与SBA的沉淀也显示出有序模式。四种类似物二元混合物的X射线晶体学数据和混合定量沉淀曲线表明,每种类似物与蛋白质都具有独特的交联晶格。与SBA显示高度组织化交联晶格的天然存在和合成碳水化合物的一个共同结构特征是,每个寡糖中存在一个假二重对称轴,该对称轴关联每个臂上的末端结合表位。这表明某些天然存在的支链寡糖的对称特征有助于与特定凝集素形成高度有序、均匀的交联复合物。