1988 Fiscal Year Final Research Report Summary
Structural Analysis of Proteoglycan Functional Domains
| Project/Area Number |
62470147
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Nagoya University |
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Principal Investigator |
SUZUKI Sakaru Nagoya Univ., Faculty of Science, Professor, 理学部, 教授 (50022504)
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| Co-Investigator(Kenkyū-buntansha) |
羽淵 弘子 名古屋大学, 理学部, 助手 (70109263)
TSUJI Masahiro Nagoya Univ., Faculty of Science, Assistant, 理学部, 助手 (80022739)
NAKANISHI Yasuo Nagoya Univ., Faculty of Science, Assistant Professor, 理学部, 助教授 (40022636)
HABUCHI Hiroko Nagoya Univ., Faculty of Science, Assistant (40022636)
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| Project Period (FY) |
1987 – 1988
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| Keywords | chondroitin sulfate proteoglycan / monoclonal antibody / fibronectin / cell-substratum adhesion / 細胞ー基質接着 / 人工プロテオグリカン |
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| Research Abstract |
1. We have previously demonstrated that chick embryo fibroblasts synthesize and secrete a large chondroitin sulfate proteoglycan (PG-M) that binds to fibronectin. We now report that chick embryo fibroblasts, BHK cells, and many other cell lines failed to adhere to fibronectin-coated substrates when PG-M was added to the medium. PG-M was also shown to inhibit the adhesion of fibroblastic cells to collagen-, or vitronection-coated substrates as well as adhesion of epithelial cells to fibronectin-coated substrates.
2. Treatment of the proteoglycan with either proteolytic enzymes or chondroitinase abolished its inhibitory effects on the fibronectin-mediated BHK cell adhesion.
3. Adhesion of the cells to laminin or GRGDS-derivatized serum albumin (RGD-containing molecules with no capacity to bind PG-M) was also inhibited by PG-M. These results indicate that the intact proteoglycan from, but not core proteinor chondroitin sulfate-free forms, is responsible for the activity, and that direct binding between PG-M and fibronectin, if any, is not a cause of the inhibition by PG-M.
4. When the immobilization of added PG-M to available plastic surfaces of fibronectin- (or GRGDS-serum albumin-) coated dishes was blocked by pretreating the dishes with serum albumin, the inhibitory effect of PG-M was abolished, suggesting that only the immobilized fraction of PG-M can act as a cell adhesion inhibitor. In immobilized form, both cartilage chondroitin sulfate proteoglycan (PG-H) and chondroitin sulfate-derivatized serum albumin also inhibited cell adhesion. In contrast, heparan sulfate proteoglycan form LD and heparan sulfate-derivatized serum albumin had far lower inhibitory activities, indicating that the active site for the interaction between cells and PG-M is on the chondroitin sulfate chain.
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