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

我们已经分离到一个编码2,610个核苷酸转录本的人ryudocan核心蛋白的cDNA，该转录本编码一个198个氨基酸的蛋白质。人和大鼠Ryudocan的核心蛋白的比较表明，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NH2和COOH末端区域，这可能在Ryudocan的生物学功能中发挥重要作用。在所有被测试的组织中都检测到一个2.7kb的主要转录本，在肺、肝、骨骼肌和肾脏的mRNA中观察到相对高水平的表达。在一些组织中也观察到了一条1.9kb的小转录本，这可能是由交替的多聚腺苷作用引起的。人ryudocan基因经荧光原位杂交定位于染色体20q12。用抗人ryudocan核心蛋白的多克隆抗体进行免疫组织化学分析，发现ryudocan在胎盘绒毛滋养层细胞和新生血管内皮细胞中均有表达，而在正常血管中不表达。人Ryudocan核心蛋白基因的分离为我们研究Ryudocan的表达调控及该分子在人内皮细胞功能中的作用奠定了基础

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