【作者】 王振新； 刘震； 高晶清；

【Author】 Zhenxin Wang;Jingqing Gao;Zhen Liu;State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences;

【机构】 中国科学院长春应用化学研究所电分析国家重点实验室；

【摘要】 <正>糖类(carbohydrates)化合物与相应的蛋白质(如lectins等)之间的相互作用广泛存在于各种生物进程(如,细胞间信息传递,细胞增值和差异化以及肿瘤细胞的转移,免疫响应等)中并起到重要的作用,其中以糖类为媒介的细胞与细胞间的识别作用是生物分子识别的重要组成,并对不同组织细胞功能分化和有序发更多还原

【Abstract】 In the post-genomic era, glycomics(the functional study of carbohydrates in living organisms) has come into the forefront of biological research because the interactions of glycoconjugates with proteins not only occur widely in biological processes of cells, including cell-cell communication, cell adhesion, fertilization, differentiation, development, inflammation, and tumor cell metastasis, but also initiate infection of host cells by bacteria and viruses. Therefore, glycoconjugates-protein interactions have been investigated by various techniques, such as surface plasmon resonance(SPR), affinity chromatography/capillary electrophoresis, quartz crystal microbalance(QCM) and carbohydrate microarray. Recently, more and more attention has been given to development of microarray-based high-throughput methods to probe the carbohydrate-protein interactions because DNA microarrays and protein microarrays have been successfully fabricated and utilized in genomic, transcriptomic, and proteomic investigations. Currently, the substrates used in carbohydrate microarrays are mainly two-dimensional(2D) planar slides which are chemical modified by a uniform monolayer of linear molecules with functional groups, such as N-hydroxysuccinimide ester(NHS), aldehyde, epoxy, and so forth. Here, a three-dimensional(3D) carbohydrate microarray-based assay has been established for studying carbohydrate-lectin binding with highly selectively. The 3D carbohydrate microarray is fabricated by immobilizing amino modified carbohydrates on the home-made porous silicon slide. The porous silicon slide was generated by electrochemical corrosion of p-type silicon wafer under acidic condition. The silylation reagents were employed as linkers for attaching the carbohydrate on the porous silicon slide. The well defined recognition systems: three monosaccharides(Man-α, Glc-β and Gal-β) with two lectins(Con A and RCA120), were chosen here to establish the microarray-based assay, respectively. Quantitative determination of the surface dissociation constants(KD,surf) for surface carbohydrates and lectins has been achieved. In addition, inhibition values(i.e., the inhibition constant(Ki)) for inhibitors in solution are also demonstrated by the saccharide competing assays.更多还原