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

我们已经分离到一系列编码2,610个核苷酸转录本的重叠的cDNA克隆，该转录本可能编码一个198个氨基酸的蛋白质，预测的相对分子质量为21,641道尔顿，用于人类Ryudocan核心蛋白。推测的人和大鼠Ryudocan核心蛋白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端，包括跨膜/细胞质结构域，其所有4个酪氨酸基的位置都是保守的，以及3个保守的糖胺多糖链附着区，这可能在Ryudocan的生物学功能中起着重要的作用。在所有被测试的组织中都检测到一个2.7kb的大转录本，在肺、肝、骨骼肌和肾脏的mRNA中观察到相对较高的表达水平。在一些组织中也观察到了一个1.9kb的小转录本，这可能是由交替的多聚腺苷作用引起的。利用荧光原位杂交技术，将人ryudocan基因定位于染色体20q12。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 series of overlapping cDNA clones encoding a 2,610 bp transcript, which potentially codes for a 198 amino acid protein with predicted molecular mass of 21,641 daltons, for the human ryudocan core protein. The deduced core proteins of the human and the rat ryudocan have high structural conservation, particularly in the NH_2 and COOH terminus regions of the putative mature core protein, including the combined transmembrane/cytoplasmic domains with conserved positions of all 4 tyrosine groups and 3 conserved glycosaminoglycan chain attachment regions, which might serve important roles for biological function of ryudocan. A major 2.7 kb 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 kb transcript was also observed in some of tissues, which would be caused by alternative polyadenylation. Human ryudocan gene has been lacalized on the chromosome 20q12 by fluorescence in situ hybridization. Isolation of the human ryudocan core protein cDNA will allow us to study for the regulation of ryudocan expression and role of this molecule in human endothelial cell function.