Spillmann D, Hård K, Thomas-Oates J, Vliegenthart J F, Misevic G, Burger M M, Finne J
Department of Medical Biochemistry, University of Turku, Finland.
J Biol Chem. 1993 Jun 25;268(18):13378-87.
The species-specific Ca(2+)-dependent reaggregation of dissociated cells of the marine sponge Microciona prolifera is mediated by a large extracellular adhesion proteoglycan. The glycans of this molecule are involved in the interactions of the proteoglycan with itself and with the sponge cells. Monoclonal antibodies against the glycans block the aggregation of sponge cells (Misevic, G. N., Finne, J., and Burger, M. M. (1987) J. Biol. Chem. 262, 5870-5877). Proteoglycan oligosaccharides were prepared by partial acid hydrolysis of the isolated glycans, and their reactivity with the monoclonal antibodies was monitored after linkage to phospholipid and immunostaining of thin layer chromatograms. One major antibody-reactive oligosaccharide was detected and purified by ion-exchange chromatography and high performance liquid chromatography. 1H NMR spectroscopy, fast atom bombardment-mass spectrometry, methylation analysis, and sequential chemical and enzymatic degradation studies indicated the structure [formula: see text] for the oligosaccharide. The depyruvylated derivative of the oligosaccharide did not react with the aggregation-blocking antibody, which indicates that the pyruvate acetal is an essential part of the epitope.
海洋海绵微拟球藻解离细胞的物种特异性钙依赖再聚集由一种大型细胞外黏附蛋白聚糖介导。该分子的聚糖参与蛋白聚糖自身以及与海绵细胞的相互作用。针对聚糖的单克隆抗体可阻断海绵细胞的聚集（米塞维克，G. N.，芬内，J.，和伯格，M. M.（1987年）《生物化学杂志》262卷，5870 - 5877页）。通过对分离出的聚糖进行部分酸水解制备蛋白聚糖寡糖，并在将其与磷脂连接及对薄层色谱进行免疫染色后监测其与单克隆抗体的反应性。通过离子交换色谱和高效液相色谱检测并纯化出一种主要的抗体反应性寡糖。1H核磁共振光谱、快原子轰击质谱、甲基化分析以及连续的化学和酶促降解研究表明该寡糖的结构为[分子式：见原文]。该寡糖的去丙酮酸化衍生物不与聚集阻断抗体反应，这表明丙酮酸缩醛是表位的重要组成部分。
