Vieira Tuane C R G, Costa-Filho Adilson, Salgado Norma C, Allodi Silvana, Valente Ana-Paula, Nasciutti Luiz E, Silva Luiz-Claudio F
Laboratório de Tecido Conjuntivo, Hospital Universitário Clementino Fraga Filho, Brazil.
Eur J Biochem. 2004 Feb;271(4):845-54. doi: 10.1111/j.1432-1033.2004.03989.x.
Acharan sulfate, a recently discovered glycosaminoglycan isolated from Achatina fulica, has a major disaccharide repeating unit of -->4)-2-acetyl,2-deoxy-alpha-d-glucopyranose(1-->4)-2-sulfo-alpha-l-idopyranosyluronic acid (1-->, making it structurally related to both heparin and heparan sulfate. It has been suggested that this glycosaminoglycan is polydisperse, with an average molecular mass of 29 kDa and known minor disaccharide sequence variants containing unsulfated iduronic acid. Acharan sulfate was found to be located in the body of this species using alcian blue staining and it was suggested to be the main constituent of the mucus. In the present work, we provide further information on the structure and compartmental distribution of acharan sulfate in the snail body. Different populations of acharan sulfate presenting charge and/or molecular mass heterogeneities were isolated from the whole body, as well as from mucus and from the organic shell matrix. A minor glycosaminoglycan fraction susceptible to degradation by nitrous acid was also purified from the snail body, suggesting the presence of N-sulfated glycosaminoglycan molecules. In addition, we demonstrate the in vivo metabolic labeling of acharan sulfate in the snail body after a meal supplemented with [35S]free sulfate. This simple approach might be applied to the study of acharan sulfate biosynthesis. Finally, we developed histochemical assays to localize acharan sulfate in the snail body by metachromatic staining and by histoautoradiography following metabolic radiolabeling with [35S]sulfate. Our results show that acharan sulfate is widely distributed among several organs.
硫酸阿查兰聚糖是最近从非洲大蜗牛中分离出的一种糖胺聚糖,其主要二糖重复单元为→4)-2-乙酰基,2-脱氧-α-D-吡喃葡萄糖(1→4)-2-磺基-α-L-艾杜糖醛酸(1→,这使其在结构上与肝素和硫酸乙酰肝素相关。有人认为这种糖胺聚糖是多分散的,平均分子量为29 kDa,并且已知含有未硫酸化艾杜糖醛酸的次要二糖序列变体。通过阿尔辛蓝染色发现硫酸阿查兰聚糖存在于该物种的体内,并且有人认为它是黏液的主要成分。在本研究中,我们提供了关于硫酸阿查兰聚糖在蜗牛体内的结构和区室分布的进一步信息。从整个身体、黏液和有机壳基质中分离出了呈现电荷和/或分子量异质性的不同群体的硫酸阿查兰聚糖。还从蜗牛体内纯化出了一个易被亚硝酸降解的次要糖胺聚糖部分,这表明存在N-硫酸化糖胺聚糖分子。此外,我们证明了在补充了[35S]游离硫酸盐的餐后,蜗牛体内硫酸阿查兰聚糖的体内代谢标记。这种简单的方法可能适用于硫酸阿查兰聚糖生物合成的研究。最后,我们开发了组织化学分析方法,通过用[35S]硫酸盐进行代谢放射性标记后的异染染色和组织放射自显影来定位蜗牛体内的硫酸阿查兰聚糖。我们的结果表明硫酸阿查兰聚糖广泛分布于多个器官中。