Carbohydrate-carbohydrate interaction in basic cell biology

基础细胞生物学中碳水化合物-碳水化合物相互作用

• 批准号: 7098818
• 负责人: Sen-itiroh Hakomori
• 金额: $ 21.36万
• 依托单位: PACIFIC NORTHWEST RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7098818
• 关键词: RNA interference; atomic force microscopy; carbohydrates; cell adhesion; cell biology; electrospray ionization mass spectrometry; enzyme linked immunosorbent assay; glycoproteins; glycosphingolipids; intermolecular interaction; laboratory mouse; lectin; nuclear magnetic resonance spectroscopy; protein binding; surface plasmon resonance; thin layer chromatography; western blottings

DESCRIPTION (provided by applicant): The concept of carbohydrate-to-carbohydrate interaction (CCI) as a basis of cell adhesion was originally developed during (i) our studies on mechanism of autoaggregation of mouse teratocarcinoma F9 cells, which mimics compaction of pre-implantation embryo; and (ii) studies by Max Burger and his group on species-specific autoaggregation of marine sponge cells. During the past decade, an increasing variety of structures have been found to be involved in either homotypic or heterotypic CCI, occurring through clusters of glycosphingolipids (GSLs) at the surfaces of two interfacing cells. CCI has been well elaborated by advanced technology in surface plasmon resonance (SPR) spectroscopy and atomic force microscopy, and its binding affinity is in the same range as that of carbohydrate-to-protein interaction (CPI) and protein-to-protein interaction (PPI). CCI has unique features not found in CPI or PPI. The specific aims of the proposed studies are: 1. Explore new cell adhesion systems based on new types of CCI detected using previously untested GSLs, and PE-conjugated N-linked or O-linked oligosaccharide probes from glycoproteins: (a) CCI based on previously-untested GSL structures; (b) CCI based on N-linked or O-linked oligosaccharides, conjugated to PE, assembled as "glycosyl SAM" or "glycochip"; (c) cell adhesion system based on the newly-found CCI. 2. Elucidate and provide evidence for involvement of CCI as initial step in physiologically or pathobiologically relevant cell adhesion systems in which CCI is suggested to occur, (a) Further studies on Lex-based CCI and cadherin-based PPI in mouse teratocarcinoma F9 cell autoaggregation. (b) Other cell adhesion systems: (i) Lex-based cell-to-cell adhesion of contact-inhibited corneal epithelial cells; (ii) Gb4-dependent CCI in autoaggregation of human teratocarcinoma cells; (iii) Ley-to-H interaction mediating adhesion of human cancer cells to endothelial cells; (iv) Mechanism of melanoma cell adhesion to mouse endothelial cells, based on GM3-to-LacCer interaction. Cell adhesion through CCI observed in Project 1 or 2 will be confirmed by (i) ruling out possible involvement of carbohydrate-binding protein; (ii) application of RNAi approach; (iii) a new SPR/ "digital window" procedure.

描述（由申请人提供）：碳水化合物到碳水化合物相互作用（CCI）作为细胞粘附的基础的概念最初是在（i）（i）对小鼠Teratocarcinoma瘤F9细胞自身凝聚机制的研究期间开发的，这些研究模仿了胚胎前胚胎栓塞前的压缩； （ii）Max Burger及其小组对海洋海绵细胞的特定物种自身聚集进行了研究。在过去的十年中，发现越来越多的结构参与了同型或异型CCI，这是通过两个接口细胞表面的糖磷脂脂（GSL）簇发生的。 CCI在表面等离子体共振（SPR）光谱和原子力显微镜中的先进技术很好地阐述了，其结合亲和力与碳水化合物与蛋白质相互作用（CPI）和蛋白质与蛋白质与蛋白质相互作用（PPI）的结合亲和力（PPI）相同。 CCI具有CPI或PPI中未发现的独特功能。 拟议的研究的具体目的是：1。基于使用先前未经测试的GSL检测到的新类型的CCI，以及糖蛋白中的PE偶联的N-链接或O-链的寡糖探针：（a）CCI基于先前已有的GSL结构； （b）基于n连锁或O连接的寡糖，与PE共轭，以“糖基SAM”或“ Glycochip”组装； （C）基于新发现的CCI的细胞粘附系统。 2。阐明并提供证据表明CCI作为生理或病理学相关的细胞粘附系统的第一步，其中建议将CCI发生，（a）进一步研究基于LEX的CCI和基于Cadherin的PPI在小鼠Teratocarcinoma瘤F9细胞自身凝集中的基于LEX的CCI和基于CADHERIN的PPI。 （b）其他细胞粘附系统：（i）基于LEX的基于LEX的细胞对接触抑制角膜上皮细胞的粘附； （ii）依赖GB4依赖性CCI在人畸胎瘤细胞自身聚集中； （iii）Ley-to-H相互作用介导人类癌细胞对内皮细胞的粘附； （iv）基于GM3-TO-LACCer相互作用，黑色素瘤细胞粘附于小鼠内皮细胞的机制。 通过（i）排除可能涉及碳水化合物结合蛋白的细胞粘附，将在项目1或2中观察到的CCI粘附； （ii）RNAi方法的应用； （iii）一个新的SPR/“数字窗口”过程。

项目成果

Self-recognition of N-linked glycans with multivalent GlcNAc, determined as ceramide mimetic conjugate.

N-连接聚糖与多价 GlcNAc 的自我识别，确定为神经酰胺模拟缀合物。

• DOI: 10.1093/glycob/cwm069
• 发表时间: 2007
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: Yoon,Seon-Joo;Ikeda,Shoko;Sadilek,Martin;Hakomori,Sen-itiroh;Ishida,Hideharu;Kiso,Makoto
• 通讯作者: Kiso,Makoto

Glycosyl conjugates of biotinylated diaminopyridine applied for study of carbohydrate-to-carbohydrate interaction.

生物素化二氨基吡啶的糖基缀合物应用于研究碳水化合物与碳水化合物的相互作用。

• DOI: 10.1007/s10719-010-9304-3
• 发表时间: 2010
• 期刊: Glycoconjugate journal
• 影响因子: 3
• 作者: Utkina,Natalia;Yoon,Seon-Joo;Hakomori,Sen-Itiroh
• 通讯作者: Hakomori,Sen-Itiroh