Molecular-patholical Analysis for the Biolocical Role of Ryudocan (Endothilal Heparan Sulfate Proteoglycan) on Vasculat Damage.

Ryudocan（内皮硫酸乙酰肝素蛋白多糖）对血管损伤的生物定位作用的分子病理分析。

• 批准号: 06836009
• 负责人: KOJIMA Tetsuhito
• 金额: $ 1.34万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 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
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06836009/
• 关键词: Ryudocan; Heparan Sulfate; Proteoglycan; Immuno-histochemistry; bFGF; midkine; TFPI; Tisssue Factor Pathway Inhibitor (TFPI); ミッドカイン(MK); Tissue Factor Pathway Inhibitor(TFPI); Inhibition radioimmuno-assay

Ryudocan, a heparan sulfate proteoglycan, was isolated from human endothelial-like EAhy926 cells by the combination of ion-exchange and immuno-affinity chromatographies. Purified human ryudocan had biochemical properties similar to those of rat ryudocan isolated from microvascular endothelial cells. Human ryudocan contained only heparan sulfate (HS) glycosaminoglycan chains along with a core protein having an apparent molecular mass of 30 kDa. We evaluated the interactions between purified human ryudocan and several extracellular ligands by using a solid-phase binding assay. It was found that basic fibroblast growth factor (bFGF), midkine (MK), and tissue factor pathway inhibitor (TFPI) exhibited significant ryudocan binding. Heparitinase, but not chondroitin ABC lyase treatment, destroyed the ability of ryudocan binding to bFGF,MK,and TFPI.Heparin and HS,but not chondroitin surfate, inhibited such ryudocan binding. Thus, the HS chains of ryudocan appear to be responsible for its binding to bFGF,MK,and TFPI.The apparent dissociation constants for purified ryudokan were bFGF,0.50 nM ; MK,0.30 nM ; and TFPI,0.74 nM.Immunohistochemical analysis revealed that ryudocan was expressed in peripheral nerve tissues, fibrous connective tissues, and placental trophoblasts. These findings suggest that ryudocan may possess multiple biologic functions, such as bFGF modulation, neurite growth promotin, and anticoagulation, via HS-binding effectors in the cellular microenvironment.

采用离子交换和免疫亲和层析相结合的方法，从人内皮样EAhy926细胞中分离到硫酸肝素蛋白多糖。纯化的人琉球多糖与从微血管内皮细胞中分离的大鼠琉球多糖具有相似的生化特性。人鱼多糖仅含有硫酸肝素（HS）糖胺聚糖链和一个表观分子质量为30 kDa的核心蛋白。我们通过固相结合实验评估了纯化的人琉球聚糖和几种细胞外配体之间的相互作用。发现碱性成纤维细胞生长因子（bFGF）、midkine （MK）和组织因子途径抑制剂（TFPI）表现出显著的ryudocan结合。肝素酶，而不是软骨素ABC裂解酶，破坏了鱼多聚糖与bFGF、MK和TFPI结合的能力。肝素和HS，而不是表面软骨素，抑制这种鱼多聚糖的结合。因此，ryudo的HS链似乎与它与bFGF、MK和TFPI的结合有关。纯化后的琉球丸表观解离常数为bFGF，0.50 nM；MK,0.30 nM；TFPI为0.74 nM。免疫组化分析显示，鱼多蛋白在周围神经组织、纤维结缔组织和胎盘滋养细胞中表达。这些发现表明，鱼多糖可能通过细胞微环境中的hs结合效应物具有多种生物学功能，如调节bFGF、促进神经突生长和抗凝血等。

项目成果

T.Kojima et al.: "Ryudocan,Its Moleculav Structure and Function." Trends in Glycosci.Glycotech.7. 385-403 (1995)

T.Kojima 等人：“Ryudocan，其分子结构和功能”。

T.Kojima et al.: "Diagnosis of Hemophilia B Carriers Using Two Novel Dinucleotide Polymorphisms and Hha I RFLP of the Factor IX Gene in Japanese Subjects" Thromb Haemost. 74. 1009-1014 (1995)

T.Kojima 等人：“在日本受试者中使用两种新型二核苷酸多态性和因子 IX 基因的 Hha I RFLP 诊断 B 型血友病携带者”血栓 Haemost。

T.Kojima et al.: "A Quantitative Protein S Deficiency Associated with a Novel Neusense Mutation and Markedly Reduceed Levcls of Mutated mRNA." Thromb.Haemost.74. 590-595 (1995)

T.Kojima 等人：“与新的 Neusense 突变和突变 mRNA 水平显着降低相关的定量蛋白 S 缺乏。”

小嶋哲人 他: "ヒト血管内皮ヘパラン硫酸プロチオグリカン-Ryudocanのコア蛋白cDNAクローニングおよび生物学的機能解析" 腎臓. 17. 41-42 (1995)

Tetsuto Kojima 等人：“人血管内皮硫酸乙酰肝素硫聚糖-Ryudocan 的核心蛋白 cDNA 克隆和生物学功能分析”肾脏。 17. 41-42 (1995)

小嶋哲人 他: "第VIII因子遺伝子逆位解析の重症血友病Aにおける保因者診断、出生前診断への応用" 臨床血液. 36. 1252-1256 (1995)

Tetsuto Kojima 等人：“因子 VIII 基因倒位分析在严重 A 型血友病携带者诊断和产前诊断中的应用”《临床血液学》36. 1252-1256 (1995)。