Nergard Cecilie Sogn, Kiyohara Hiroaki, Reynolds James C, Thomas-Oates Jane E, Matsumoto Tsukasa, Yamada Haruki, Patel Trushar, Petersen Dirk, Michaelsen Terje E, Diallo Drissa, Paulsen Berit Smestad
Department of Pharmacognosy, School of Pharmacy, University of Oslo, P.O. Box 1068, Blindern, 0316 Oslo, Norway.
Biomacromolecules. 2006 Jan;7(1):71-9. doi: 10.1021/bm050355g.
Structures of three pectic arabinogalactans, one from Vernonia kotschyana (Vk2a) and two from Cochlospermum tinctorium (Ct50A1 and Ct50A2), and their complement fixation and induction of B cell proliferation in vitro were compared. The polysaccharide Vk2a expressed potent biological activity in both assays compared with Ct50A1 and Ct50A2. Vk2a possessed a very high molecular weight (1150 +/- 20 kDa) compared with Ct50A1 and Ct50A2 which both showed a polydisperse nature with the highest molecular weight polymers in each fraction estimated at approximately 105 kDa (Ct1a) and 640 +/- 100 kDa (Ct2a), respectively. The HMW polymers showed complement fixation in the same range as the native fractions. The arabinogalactan II content was low in Vk2a (2%) compared with that in Ct50A1 (23%) and Ct50A2 (12%). The high molecular weight polymers were subjected to digestion with a beta-d-(1, 3)-galactanase-rich fraction from Driselase, oligomers were isolated by HPAEC, and their finer structures were determined by MALDI- and ES-qoToF-MS, linkage, and monosaccharide composition analyses. Vk2a consists of both a galacturonan core and a rhamnogalacturonan core rich in neutral side chains. The backbones of both Ct-polysaccharides consist mainly of RG-I regions with numerous neutral side chains dominated by galactosyl residues, whereas the homogalacturonan regions seem to be small. Differences in the chain lengths of the 6-linked galacto-oligosaccharides attached to the 3-linked galactan core could not be related to the differences in the potencies of the biological activities observed.
比较了三种果胶阿拉伯半乳聚糖的结构,一种来自科奇亚斑鸠菊(Vk2a),两种来自染色柯桠(Ct50A1和Ct50A2),并比较了它们的补体固定作用以及体外诱导B细胞增殖的能力。与Ct50A1和Ct50A2相比,多糖Vk2a在这两种测定中均表现出强大的生物活性。与Ct50A1和Ct50A2相比,Vk2a具有非常高的分子量(1150±20 kDa),Ct50A1和Ct50A2均呈现多分散性质,每个级分中分子量最高的聚合物估计分别约为105 kDa(Ct1a)和640±100 kDa(Ct2a)。高分子量聚合物的补体固定作用与天然级分处于相同范围。与Ct50A1(23%)和Ct50A2(12%)相比,Vk2a中的阿拉伯半乳聚糖II含量较低(2%)。将高分子量聚合物用来自Driselase的富含β-d-(1, 3)-半乳聚糖酶的级分进行消化,通过高效阴离子交换色谱(HPAEC)分离低聚物,并通过基质辅助激光解吸电离-和电喷雾四极杆飞行时间质谱(MALDI-和ES-qoToF-MS)、连接分析和单糖组成分析确定其更精细的结构。Vk2a由一个半乳糖醛酸聚糖核心和一个富含中性侧链的鼠李糖半乳糖醛酸聚糖核心组成。两种Ct-多糖的主链主要由具有大量以半乳糖基残基为主的中性侧链的鼠李糖半乳糖醛酸聚糖-I(RG-I)区域组成,而同型半乳糖醛酸聚糖区域似乎较小。连接在3-连接的半乳聚糖核心上的6-连接的半乳糖寡糖的链长差异与所观察到的生物活性强度差异无关。