Sato Kazuki, Hara Katsuya, Yoshimi Yoshihisa, Kitazawa Kiminari, Ito Haruka, Tsumuraya Yoichi, Kotake Toshihisa
Department of Biochemistry and Molecular Biology, Faculty of Science, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama, 338-8570, Japan.
Division of Life Science, Graduate School of Science and Engineering, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama, 338-8570, Japan.
Carbohydr Res. 2018 Sep;467:8-13. doi: 10.1016/j.carres.2018.07.004. Epub 2018 Jul 7.
Larch arabinogalactan (AG) is classified as a plant type II AG. Its basic structure is constituted by a β-1,3-galactan main chain with β-1,6-galactan side chains. But its properties are distinct from other type II AGs. Whereas most type II AGs are found as glycan moieties of arabinogalactan-protein (AGP), larch AG lacks a protein moiety. Larch AG itself is also unlike other type II AGs as it lacks Yariv reactivity, the capability of AG to form insoluble precipitate with β-Yariv reagents, 1,3,5-tri-(p-glycosyloxyphenylazo)-2,4,6-trihydroxybenzene with β-glucosyl or β-galactosyl residues at the termini. For the present study, we prepared β-galactan I, II, and III from larch AG by performing single, double, and triple Smith degradation, which breaks β-1,6-galactan side chains, and examined Yariv reactivity of the products. Methylation analysis revealed that β-galactans II and III had lost more than 90% of their β-1,6-galactan branches. In the radial gel diffusion assay, β-galactans II and III showed Yariv reactivity, indicating the presence of a Yariv-reactive structure in larch AG, although native larch AG does not have Yariv reactivity. The Yariv reactivity of the β-galactans was completely lost after treatment with endo-β-1,3-galactanase. These results confirm that β-1,3-galactan is necessary for Yariv reactivity of type II AG. The present results also suggest that high substitution of β-1,3-galactan with β-1,6-galactan side chains affects Yariv reactivity in larch AG.
落叶松阿拉伯半乳聚糖(AG)被归类为植物II型AG。其基本结构由具有β-1,6-半乳聚糖侧链的β-1,3-半乳聚糖主链构成。但其性质与其他II型AG不同。大多数II型AG以阿拉伯半乳聚糖蛋白(AGP)的聚糖部分形式存在,而落叶松AG缺乏蛋白质部分。落叶松AG本身也与其他II型AG不同,因为它缺乏Yariv反应性,即AG与β-Yariv试剂(1,3,5-三-(对-糖基氧基苯基偶氮)-2,4,6-三羟基苯)在末端带有β-葡萄糖基或β-半乳糖基残基形成不溶性沉淀的能力。在本研究中,我们通过进行单、双和三次Smith降解从落叶松AG制备了β-半乳聚糖I、II和III,Smith降解会断裂β-1,6-半乳聚糖侧链,并检测了产物的Yariv反应性。甲基化分析表明,β-半乳聚糖II和III失去了超过90%的β-1,6-半乳聚糖分支。在径向凝胶扩散试验中,β-半乳聚糖II和III表现出Yariv反应性,表明落叶松AG中存在Yariv反应性结构,尽管天然落叶松AG没有Yariv反应性。用内切β-1,3-半乳聚糖酶处理后,β-半乳聚糖的Yariv反应性完全丧失。这些结果证实β-1,3-半乳聚糖对于II型AG的Yariv反应性是必需的。目前的结果还表明,β-1,3-半乳聚糖被β-1,6-半乳聚糖侧链高度取代会影响落叶松AG中的Yariv反应性。