Molecular and Pathological Analysis of Anticoagulant Heparan Sulfate Proteoglycan from Endothelial Cell

内皮细胞抗凝硫酸乙酰肝素蛋白多糖的分子和病​​理学分析

• 批准号: 05454330
• 负责人: SAITO Hidehiko
• 金额: $ 4.86万
• 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 (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454330/
• 关键词: Endothelial cell; Heparan Sulfate; Proteoglycan; Ryudocan; cDNA; Immuno-histochemistry; Inhibition radioimmuno assay; 血管内皮; alternative polyadenylation; ヒト染色体20q12

We have isolated a cDNA of the human ryudocan core protein encoding a 2,610 bp transcript, which potentially codes for a 198 amino acid protein. Comparison of the deduced core proteina between the human and the rat ryudocan revealed that they have high structural conservation, particularly in the NH_2 and COOH terminus regions of the putative mature core protein, which might serve important roles for biological function of ryudocan. A major 2.7kb transcript was detected in all tissues tested, with relatively high levels of expression observed in mRNA from lung, liver, skeletal muscle and kidney. A minor 1.9 kbtranscript was also observed in some of tissues, which would be caused by alternative polyadenylation. Human ryudocan gene has localized on the chromosome 20q12 by fluorescence in situ hybridization. Immuno-histochemical analysis using a specific polyclonal antibody against human ryudocan core protein revealed that ryudocan was expressed in trophoblasts of placental villi and in endothlial cells of neovesseles, but not in those of normal vesseles. Isolation of the human ryudocan core protein cDNA will allow us to study for the regulation of ryudocan expression and the role of this molecule in human endothelial cell function

我们已经分离了编码2，610 bp转录本的人ryudocan核心蛋白的cDNA，其可能编码198个氨基酸的蛋白。比较人和大鼠ryudocan的核心蛋白，发现它们具有高度的结构保守性，特别是在推测的成熟核心蛋白的NH_2和COOH末端区域，这可能对ryudocan的生物学功能起重要作用。在所有测试的组织中检测到主要的2.7kb转录物，在来自肺、肝、骨骼肌和肾的mRNA中观察到相对高水平的表达。在某些组织中也观察到少量的1.9kb转录本，这可能是由选择性多聚腺苷酸化引起的。用荧光原位杂交技术将人ryudocan基因定位于染色体20q12。用抗人ryudocan核心蛋白的特异性多克隆抗体进行免疫组化分析，发现ryudocan在胎盘绒毛滋养层细胞和新生血管内皮细胞中表达，而在正常血管中不表达。人ryudocan核心蛋白cDNA的分离将使我们能够研究ryudocan表达的调控及其在人内皮细胞功能中的作用

项目成果

Kojima,T.: "Human ryudocan core protein:molecular cloning and charcterization of the cDNA,and chromosomal localization of the gene." Biochem Bioph Res Comm. 190. 814-822 (1993)

Kojima,T.：“人类龙道坎核心蛋白：cDNA 的分子克隆和表征，以及基因的染色体定位。”

Yamamoto,K.: "Homozygous protein C deficiency:indentification of a novel missense mutation that causes impaired secretion of the mutant protein C." J Lab Clin Med. 119. 87-95 (1992)

Yamamoto,K.：“纯合蛋白 C 缺陷：鉴定出一种新的错义突变，该突变会导致突变蛋白 C 的分泌受损。”

Sugiura,I.: "Three distinct point mutations of the von Willebrand factor gene in four patients with type IIA von Willebrand disease." Thromb Haemost. 67. 612-617 (1992)

Sugiura,I.：“四名 IIA 型冯维勒布兰德病患者的冯维勒布兰德因子基因出现三种不同的点突变。”

Hidehiko Saito et al.: "Human Ryudocan Core Protein:Molecular Cloning and Chavacteriqation of the cDNA,and Chromosomal Localiqation of the Gene." Biochem.Biophys.Res.Commun.190. 814-822 (1993)

Hidehiko Saito 等人：“人类 Ryudocan 核心蛋白：cDNA 的分子克隆和 Chavacteriqation，以及基因的染色体定位。”

Hidehiko Saito et al.: "Human Ryudocan Core Protein : Molecular Cloning and Charaterization of the cDNA,and Chromosomal Localization of the Gene" Biochem.Biophys.Res.Commun. 190. 814-822 (1993)

Hidehiko Saito 等人：“人类 Ryudocan 核心蛋白：cDNA 的分子克隆和表征，以及基因的染色体定位”Biochem.Biophys.Res.Commun。