利用糖芯片研究半乳寡糖与蓖麻凝集素和鸡冠刺桐凝集素间的相互作用  

Investigation of Interaction of Galacto-oligosaccharides with Ricinus Communis Agglutinin 120 and Erythrina Cristagalli Lectin by Glycochip

作　　者：王玉峰[1,2] 吴建东[1,3] 韩章润[1,3] 吕友晶[1,3] 赵峡[1,3] 于广利[1,3] 

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,青岛266003 [2]江苏雨润肉类产业集团有限公司,南京210041 [3]中国海洋大学山东省糖科学与糖工程重点实验室,青岛266003

出　　处：《分析化学》2012年第9期1341-1346,共6页Chinese Journal of Analytical Chemistry

基　　金：科技部国际合作项目(No.2007DFA30980);国家自然科学基金(Nos.31070724,91129706);海洋公益专项(No.201005024);长江学者与创新团队项目(No.IRT0944)资助

摘　　要：建立了用糖芯片技术研究寡糖与凝集素相互作用的方法。以新乳糖-N-四糖(LNnT)、乳糖-N-四糖(LNT)、λ-卡拉胶四糖(L4)和琼胶五糖(A5)为原料,经还原胺化法分别将其与1,2-十六烷基磷脂酰乙醇胺偶联得到拟糖脂,再将其与卵磷脂和胆固醇按4∶2∶5比例混合制备成脂质体后,用全自动芯片点样仪将其点印在硝酸纤维素膜包被的玻片上制成糖芯片,并进行寡糖与凝集素的结合实验。结果表明,蓖麻凝集素(Ricinus communis agglutinin 120,RCA120)特异性识别非还原端糖残基为Galβ(1#4)的寡糖,而鸡冠刺桐凝集素(Erythrina cristagalli lectin,ECL)特异性识别非还原端糖残基为Galβ(1#4)GlcNAc的LNnT。采用接触式芯片点样仪制备糖芯片,并用荧光扫描仪对糖与凝集素结合信号进行检测,增加了灵敏度和准确性。本方法不仅适合于糖与蛋白相互作用的研究,也有助于加快糖类药物的发现。A method was established to study the interaction between lectins and galacto oligosaccha- rides by glycochip technology. Four oligosaccharides Lacto-neo N-tetraose （LNnT）, Lacto N- tetraose （LNT）, 2 carra-tetrasaccharide （L4） and agaro-pentosaccharide （AS）, were converted to neoglycolipids （NGLs） by conjugation to 1,2 dihexadecyl sn-glycero-3-phosphoethanolamine （DHPE） via reductive animation. Liposome was prepared with the ratio of 4：2：5 of NGLs： phosphatidylcho- line： cholesterol, respectively. The NGL liposome was printed onto the nitrocellulose membranes coated glass plate to get glycochips, and the interaction of oligosaccharides with lectins was investi- galed The results demonstrated that Ricinus communis agglutinin 120 （RCA,20） bound to the oligo- saccharides whose non-reducing terminal was Gal β（1→4）, and Erythrina cristagalli lectin （ECL） only bind with LNnT whose non-reducing terminals were Gal 13（1→4）GlcNAc. The preparation method of glycochip by contactive microarrayer was established, the binding signals of oligosaccharides with lectins were analyzed by fluorescence scanner, and the sensitivity and accuracy were dramatically enhanced. This method is not only suitable for the interaction study between oligosaccharides and proteins, but can also speed up the carbohydrate-based drug discovery.

关 键 词：糖芯片 半乳寡糖 拟糖脂 凝集素 

分 类 号：R284.1[医药卫生—中药学]