Initiation Mechanism of Extrinsic Blood Coagulation System

外源性凝血系统的启动机制

• 批准号: 63044110
• 负责人: IWANAGA Sadaaki
• 金额: $ 5.76万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63044110/
• 关键词: Factor VII; EGF-like domain; Oligosaccharide; Protein Z; Serine protease; cDNA cloning; Factor IX; gamma-Carboxyglutamic acid residue

The research progress of this year was as follows :1. We reported the presence of a new trisaccharide composed of two xylose and reducing terminal glucose residues linked to serine residues of bovine blood clotting factors VII and IX (Hase, S., Kawabata, S., Nishimura H., Takeya, H., Sueyoshi, T., Miyata, T., Iwanaga, S., Takao, T., Shimonishi, Y., and Ikenaka, T. (1988) J. Biochem. (Tokyo) 104, 867-868). We describe here the detailed structural analysis of the trisaccharide. Glycopeptides were prepared from bovine factor IX by digestion with Pronase followed by purification by column chromatography. The trisaccharide was released from the protein by the beta-elimination reaction with hydrazine, and the reducing end of the sugar chain was tagged with 2-aminopyridine. The fluorescent pyridylamino derivative of the trisaccharide was purified by gel filtration and reversed-phase high performance liquid chromatography. The glycopeptides and pyridylamino-trisaccharide thus obtained were sub … More jected to methylation study. 500-MHz ^1H nuclear magnetic resonance spectroscopy, and periodate oxidation. Glucose and xylose belong to the D series by high performance liquid chromatography on a chiral column. From the results, the structure of the trisaccharide is proposed as : D-Xylpalpha1-3-D-Xylpalpha-1-3-D-Glcpbeta1-O-Ser-53.2. Protein Z is a vitamin K-dependent glycoprotein isolated and characterized from human and bovine plasma. A cDNA coding for human protein Z has been obtained by the isolation of phage clones from a liver cDNA library and in vivo amplification of two other liver libraries. Protein Z is synthesized with a prepro-leader sequence of 40 amino acids. The mature protein is composed of 360 residues including a Gla domain of 13 carboxyglutamic acid residues, two epidermal growth factor domains, and a carboxyl terminal region which is highly homologous to the catalytic domain of serine proteases. Human protein Z, however, contains an Asp instead of Ser and a Lys instead of His in the catalytic triad of the active site. Less

今年的研究进展如下：1。我们报道了由两个木糖和还原末端葡萄糖与牛血液凝结因子VII和IX相关的新三糖的存在（Hase，S.，Kawabata，S.，Nishimura H.，Nishimura H.，Takeya，Takeya，H. Shimonishi，Y。和Ikenaka，T。（1988）J。Biochem（东京）104，867-868）。我们在这里描述了三糖的详细结构分析。糖肽是通过植物酶消化从牛IX中制备的，然后通过柱色谱纯化。三糖通过与氢嗪的β-脱酮反应从蛋白质中释放出来，并用2-氨基吡啶标记了糖链的还原端。通过凝胶过滤和反相高性能液相色谱纯化三糖的荧光吡啶基氨基衍生物。因此获得的糖肽和吡啶基氨基三糖含量为sub…更受甲基化研究。 500-MHz ^1H核磁共振光谱和周期氧化物。葡萄糖和木糖属于手性柱上的高性能液相色谱。从结果中，提出了三糖的结构：d-xylpalpha1-3-d-d-xylpalpha-1-3-d-d-d-d-d-d-d-glcpbeta1-o-ser-53.2。蛋白Z是一种从人和牛等离子体中分离出来的维生素K依赖性糖蛋白。通过从肝cDNA文库中分离噬菌体克隆和其他两个肝脏库的体内扩增，已经获得了编码人蛋白Z的cDNA。蛋白Z与40个氨基酸的前领导者序列合成。成熟的蛋白质由360个残基组成，包括13个羧酸谷氨酸保留的GLA结构域，两个表皮生长因子结构域和一个与丝氨酸蛋白催化结构域高度同源的羧基末端区域。但是，人蛋白Z包含一个ASP，而不是SER和LYS，而不是在活性位点的催化三合会中。较少的

项目成果

Muta, T. et al.: "Tachyplesins Isolated from Hemocytes of South Asian Horseshoe Crabs (Carcinoscorpius rotundicauda and Tachypleus gigas) : Identification of a New Tachyplesin, Tachyplesin III. and a Processing Intermediates of Its Precursor." J. Biochem.

Muta, T. 等人：“从南亚鲎（Carcinoscorpius rotundicauda 和 Tachypleus gigas）血细胞中分离出的鲎素：鉴定一种新的鲎素，鲎素 III 及其前体的加工中间体。”

Takeya,H.et al.: "The Complete Amino Acid Sepuence of the High Molecular Mass Hemorrhagic Protein,HR1B,Isolated from the Venom of Trimeresurus flavoviridis." J.Biol.Chem.265. 16068-16073 (1990)

Takeya,H.et al.：“从竹叶青竹叶青毒液中分离出的高分子出血蛋白 HR1B 的完整氨基酸序列。”

Niwa, M. et al.: "Biological Activities of Anti-LPS Factor and LPS Binding Peptide From Horseshoe Crab Amebocytes." Adv. Exp. Med. Biol.256. 257-271 (1990)

Niwa, M. 等人：“来自鲎变形细胞的抗 LPS 因子和 LPS 结合肽的生物活性”。

Ichinose, A. et al.: "Amino Acid Sequence of Human Protein Z, a Vitamin K-dependent Plasma Glycoprotein." Biochem. Biophys. Res. Communs.172. 1139-1144 (1990)

Ichinose, A. 等人：“人类 Z 蛋白的氨基酸序列，一种维生素 K 依赖性血浆糖蛋白。”

Kumatori,A.et al.: "cDNA Cloning and Sequencing of Component C9 of Proteosomes from Rat Hepatoma Cells." FEBS Lett.264. 279-282 (1990)

Kumatori,A.et al.：“大鼠肝癌细胞蛋白体 C9 组分的 cDNA 克隆和测序”。