1987 Fiscal Year Final Research Report Summary
Isolation of Endothelial Cell Proteoheparan Sulfate and Preparationof Its Antibody
| Project/Area Number |
61570420
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Kochi Medical School |
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Principal Investigator |
SHIMADA Kazuyuki Kochi Medical School, 医学部, 講師 (90145128)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUBAYASHI Kozo Kochi Medical School, 医学部, 助手 (70190494)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Endothelial cell / Heparan sulfate / Chondroitin sulfate / <beta>-D-Xyloside / n-butyrate / アンチトロンビンIII |
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| Research Abstract |
In the course of isolation of endothelial cell heparan sulfate, various agents (<beta>-D-xyloside, n-butyrate) were found to alter the property of cell-associated proteoheparan sulfate. Incubation of endothelial cell cultures with <beta>-D-xyloside resulted in a parallel reduction of antithrombin III binding and biosynthesis of cell surface heparan sulfate. N-butyrate caused the increase in the binding, but was cytotoxic. <beta>-D-xyloside did not affect the cellular growth. Whereas the size of cell surface heparan sulfate was not altered by xyloside treatment, it apperaed to have slightly less net negative charge and a significantly reduced proportion of the molecule with high affinity for antithrombin III in the presence of xyloside. On the other hand, secretion of chondroitin sulfate chains into the medium was markedly increased in the presence of xyloside, accompanied by a smaller increase in secretion of free heparan sulfate chains. These results suggest that <beta>-D-xyloside caused a decrease in production as well as some subtle structural alterations of cell-surface-associated heparan sulfate, which could serve as binding sites for antithrombin III. Furthermore, more importantly increased amounts of heparan sulfate in the medium from xyoside-treated cells suggest that this system may provide a potentially useful source for the isolation of this compound. We are continuing this project in this direction.
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