REGULATION OF VARIOUS CELL BEHAVIORS BY PROTEOGLYCANS THAT INHIBIT CELL-ADHESION,ANTI-ADHESIVE MOLECULES

抑制细胞粘附、抗粘附分子的蛋白聚糖对多种细胞行为的调节

基本信息

批准号：
06454647
负责人：
KIMATA Koji
金额：
$ 4.03万
依托单位：
INSTITUTE FOR MOLECULAR SCIENCE OF MEDICINE,AICHI MEDICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

1) Studies on the mechanisms for nati-adhesive activity of PG-M : In order to suggest a molecular biological strategy for glycocalfin which is a cell-surface receptor postulated to transmit the antiadhesive activity of PG-M to cells we first tried to isolate glycocalfin cDNA.Partial amino acid sequences and the immunoreactivity to various antibodies of glycocalfin purified from cultured human cells by the affinity chromatography using chondroitin sulfate-conjugated gels have revealed the complete identity of glycocalfin to annexin VI.This conclusion has enabled us to obtain the cDNA,because the cDNA is already available. Further studies on the anti-adhesive activity of PG-M using the following three different types of assay methods have convinced the above strategy : Comparisons of cell adhesion to fibronectin-coated U-shape dishes under the centrifugal force ; binding activities of liposome-intercalated integrin to substrates ; adhesion of half-denatured cells to substrates. All the results have suggested that PG-M only interrupt cell-spreading but not cell-binding probably by glycocalfin-mediated signal transduction. (2) Regulation of cell behaviors by anti-adhesion activity of PG-M : We have finished the cDNA analysis, northern analysis, and genomic DNA analysis for chicken, mouse, and human PG-Ms. The results have suggested the presence of multiple forms of PG-M with different anti-adhesive activity due to the different chondroitin sulfate chain number and their tissue- and developmental stage-dependent alterations. For example, the largest form of PG-M (the richest in the chain) was found in the early stages of development. It is likely, therefore, that specific mechanisms may be involved in the anti-adhesion of PG-M.

1) PG-M 抗粘附活性机制的研究：为了提出糖钙蛋白的分子生物学策略，糖钙蛋白是一种细胞表面受体，假定将 PG-M 的抗粘附活性传递给细胞，我们首先尝试分离糖钙蛋白 cDNA。通过使用硫酸软骨素缀合凝胶的亲和层析从培养的人细胞中纯化的糖钙蛋白的部分氨基酸序列和对各种抗体的免疫反应性已揭示糖钙蛋白与膜联蛋白VI的完整同一性。这个结论使我们能够获得cDNA，因为cDNA已经可用。使用以下三种不同类型的测定方法对 PG-M 的抗粘附活性的进一步研究证实了上述策略：离心力下细胞对纤连蛋白包被的 U 形培养皿的粘附的比较；脂质体嵌入的整合素与底物的结合活性；半变性细胞与基质的粘附。所有结果表明，PG-M 可能通过糖钙蛋白介导的信号转导仅中断细胞扩散，但不中断细胞结合。 (2) PG-M的抗粘附活性对细胞行为的调节：我们已经完成了鸡、小鼠和人PG-M的cDNA分析、Northern分析和基因组DNA分析。结果表明，由于不同的硫酸软骨素链数及其组织和发育阶段依赖性改变，存在具有不同抗粘连活性的多种形式的 PG-M。例如，PG-M的最大形式（链中最丰富）是在开发的早期阶段发现的。因此，PG-M 的抗粘连可能涉及特定机制。