MODULATION OF CELL BEHAVIORS BY PROTEOGLYCANS WITH ANTI-CELL-SUBSTRATE ADHESION ACTIVITY-STUDIES ON INVOLVING MOLECULES AND THE MECHANISM

具有抗细胞基质粘附活性的蛋白多糖对细胞行为的调节作用-涉及分子及机制的研究

基本信息

批准号：
04454595
负责人：
KIMATA Koji
金额：
$ 3.33万
依托单位：
INSTITUTION 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：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454595/
关键词：
cell adhesion anti-adhesion proteglycans chondroitin osteosacoma PG-M glycocalfin annexin 58プロテイン軟骨分化

项目摘要

Cell adhesion to substrate or cell is a crucial step that regulates a variety of sell behavior. We previously showed that PG-M, a large chondroitin sulfate proteoglycan had an jnhibitory activity for any types of cell-adhesion to ECM and proposed its general role in modulating sell-ECM interactions (J.Biol. Chem., 264, 8012, 1989) Several lines of evidence suggested that the activity could be due to the chondroitin sulfate chains immobilized onto ECM via the core protein moiety. To obtain the direct evidence for the role of PG-M, we investigated the effect of the anitisense specific inhibition of PG-M synthesis on the oncogenic adhesion of human osteosarcoma cells. The inhibition suppressed the malignant cell-adhesive phenotype, consistent with the idea that PG-M controls sell-ECM adhesion. To gain insight into the mechanism, we postulated that the chondroitin sulfate-induced inhibition of sell adhesion might be due to a sell surface receptor capable of intercing specifically with chondroitin sulfate chains, thereby influencing the clustering or conformational change of receptors for ECM molecules. The present results have indicated the occurrence of a 58-kDa protein which had an affinity to the PG-M- OR chondroitin sulfate-immobilized gel column. The interaction of the protein with the column required the presence of Ca^<2+>. According to these properties, 58-kDa protein is now designated glycocalfin could be extracted with a detergent-contergent-containing solution from the membrane fractions of cultured fibroblasts and from the homogenate of minced embryo bodirs. The amino asid sequences of purified of gycocolfin have shown that this molecule is a famiry of annexin VI.Further studies of the protein will answer the question whether our postulated mechanism for the anti-adhesion actibity of immobilized chondroitin sulfate chains could be operative or not.

细胞对基材或细胞的粘附是调节各种销售行为的关键步骤。我们先前表明，PG-M是一种大型硫酸软骨素蛋白聚糖，对任何类型的ECM细胞粘附具有JNHIBNOITION活性，并提出了其在调节卖出ECM相互作用中的一般作用（J.Biol。Chem。，264，8012，1989 1999 ）几条证据表明，该活性可能是由于硫酸软骨链通过核心蛋白部分固定在ECM上的原因。为了获得PG-M作用的直接证据，我们研究了PG-M合成对人骨骨肉瘤细胞致癌粘附的影响。抑制作用抑制了恶性细胞粘附表型，这与PG-M控制卖出ECM粘附的想法一致。为了深入了解该机制，我们假设硫酸软骨蛋白诱导的销售粘附抑制可能是由于销售表面受体能够特异性地与硫酸软骨软骨链介导，从而影响了ECM分子受体的聚类或构象变化。目前的结果表明，发生了58 kDa蛋白，该蛋白与硫酸盐或软骨素硫酸盐含量的凝胶柱具有亲和力。蛋白质与色谱柱的相互作用需要Ca^<2+>的存在。根据这些特性，现在可以从培养的成纤维细胞的膜馏分和切碎的胚胎Bodirs的均匀材料中提取58 kDA蛋白的糖蛋白。 Gycocolfin纯化的氨基ASID序列表明，该分子是蛋白质膜毒素VI的著名研究。该蛋白质的研究将回答我们假定的抗粘附活性机制是否可以操作。