从海贻贝Crenomytilus grayanus中分离并鉴定新型N-乙酰半乳糖胺/半乳糖特异性凝集素

Isolation and characterization of new GalNAc/Gal-specific lectin from the sea mussel Crenomytilus grayanus.

作者信息

Belogortseva N I, Molchanova V I, Kurika A V, Skobun A S, Glazkova V E

机构信息

Pacific Institute of Bioorganic Chemistry, Far East Branch, Russian Academy of Science, Vladivostok, Russia.

出版信息

Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1998 Jan;119(1):45-50. doi: 10.1016/s0742-8413(97)00180-1.

DOI:10.1016/s0742-8413(97)00180-1

PMID:9568372

Abstract

A lectin, Crenomytilus grayanus (CGL), was purified from sea mussel C. grayanus by affinity chromatography on acid-treated Sepharose 6B and following gel filtration on Sephacryl S-200. Molecular weight of the lectin obtained was determined by SDS-PAGE to be 18,000, independent of the presence or absence of beta-mercaptoethanol. CGL was found to agglutinate all types of the human erythrocytes together with mouse and rabbit. In hemagglutination inhibition assays, N-acetyl-D-galactosamine, D-galactose, and D-talose were the most potent inhibitors among the monosaccharides tested. Out of the oligosaccharides containing nonreducing terminal D-galactose, melibiose, and raffinose were found to be strong inhibitors. On the other hand, among the glycoproteins, asialo-BSM was the best inhibitor. The hemagglutinating activity of CGL was independent of the divalent cations Ca2+ and Mg2+. Significant CGL activity was observed between pH 8-10.

摘要

从海贻贝（Crenomytilus grayanus）中通过在酸处理的琼脂糖凝胶6B上进行亲和层析以及随后在Sephacryl S - 200上进行凝胶过滤，纯化得到一种凝集素，即灰贻贝凝集素（CGL）。通过SDS - PAGE测定所得凝集素的分子量为18,000，与β-巯基乙醇的存在与否无关。发现CGL能凝集所有类型的人红细胞以及小鼠和兔的红细胞。在血凝抑制试验中，N - 乙酰 - D - 半乳糖胺、D - 半乳糖和D - 塔罗糖是所测试单糖中最有效的抑制剂。在含有非还原末端D - 半乳糖的寡糖中，蜜二糖和棉子糖是强抑制剂。另一方面，在糖蛋白中，去唾液酸牛血清白蛋白是最佳抑制剂。CGL的血凝活性与二价阳离子Ca2 +和Mg2 +无关。在pH 8 - 10之间观察到显著的CGL活性。