Structural analysis of oligosaccharides ligands to a serum lectin inducing an anti-tumor activity

诱导抗肿瘤活性的血清凝集素寡糖配体的结构分析

基本信息

批准号：
14572054
负责人：
KAWASAKI Nobuko
金额：
$ 2.5万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572054/
关键词：
serum lectin mannan-binding protein mannose-binding protein human colon cancer cell anti-tumor activity host defence factor oligosaccharide ligand polylactosamine structure ヒト結腸ガン細胞赤血球Band3糖タンパク質

项目摘要

Serum Mannan-binding protein (MBP), a C-type mammalian lectin specific for mannose, N-acetylglucosamine and fucose, has a potent growth inhibitory activity to a human colorectal carcinoma cell line in vivo. In this study, we have characterized the MBP-ligand oligosaccharides expressed on the human colorectal carcinoma cell line, SW 1116, which are suggested to trigger anti-tumor activity of MBP.1.FITC-labeled MBP binding to SW1116 cells was inhibited specifically by the ligand sugars, a fucose-specific lectin, and anti-Lewis A and anti-Lewis B mAbs, suggesting that MBP binds to type-I carbohydrates.2.MBP-ligand glycopeptides were isolated by a MBP affinity column from the high molecular weight glycopeptides fraction of the pronase-digest of SW 1116 cell lysates. After hydrazinolysis followed by pyridylamination of MBP-ligand glycopeptides, PA-derivatized MBP-ligand oligosaccharides were isolated again by the MBP affinity column.(1)Carbohydrate analysis indicated that MBP-ligand oligosaccharides contained high molecular size N-glycans with high galactose, N-acetylglucosamine and fucose contents.(2)Endo-β-galactosidase digestion of the MBP-ligand oligosaccharides resulted in a marked reduction of the binding activity to the MBP column together with the decrease of their molecular sizes, indicating the presence of poly N-acetyllactosamine structures. The selective removal of fucose residues from MBP-ligand oligosaccharides resulted in almost complete loss of the binding activity to the MBP and the AAL affinity columns.(3)MALDI-MSIMS and nano ESI-MS/MS analyses of the MBP-ligand oligosaccharides indicated the presence of highly fucosylated structure.MBP-ligand oligosaccharides appear to be high molecular weight poly N-acetyllactosamine-type with high fucose content.

血清Mannan结合蛋白（MBP）是一种针对甘露糖，N-乙酰葡萄糖和富谷糖的C型哺乳动物讲座，具有对人体内有色癌细胞系的潜在生长抑制活性。在这项研究中，我们表征了在人类有色癌细胞系SW 1116上表达的MBP - 配体寡糖，建议触发MBP.1.FITC标记的MBP MBP的抗肿瘤活性。 mAb，表明MBP与I型碳氢化物结合。2.Mbp-cligand糖肽通过MBP亲和力柱从高分子量糖肽分数的SW 1116细胞裂解液的基础酶数分数中分离出来。 After hydrozinolysis was followed by pyridylation of MBP-ligand glycopeptides, PA-derivatized MBP-ligand oligosaccharides were isolated again by the MBP affinity column.(1)Carbohydrate analysis indicated that MBP-ligand oligosaccharides contained high molecular size N-glycans with high galactose, N-acetylglucosamine and fucose （2）MBP - 配体寡糖的内部-β-半乳糖苷酶消化导致与MBP柱的结合活性显着减少，并减少其分子大小，表明选择性去除MBP-LigosAcAcAcac的葡萄糖残差几乎导致MBP寡糖的损失，几乎导致了一定的活性。（3）MALDI-MSIMS和纳米ESI-MS/MS/MS/MS/MS分析MBP-ligans寡糖表明存在高度诱导的结构。MBP-ligosAcacacyAcachiese。MBP-LigosAcacacy似乎是高分子寡糖，是高分子量的多聚乙酰氨基甲胺氨基氨基氨基氨基胺含量，具有较高的葡萄糖含量。