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

本年度的研究进展如下：1.我们报道了一种新的三糖的存在，该三糖由两个木糖和连接到牛凝血因子VII和IX的丝氨酸残基的还原性末端葡萄糖残基组成（Hase，S.，Kawabata，S.，西村H.，Takeya，H.，末吉，T.，宫田，T.，Iwanaga，S.，Takao，T.，Shimonishi，Y.，和Ikenaka，T.（Tokyo）104，867-868）。我们在这里描述的三糖的详细结构分析。通过链霉蛋白酶消化，然后通过柱层析纯化，从牛因子IX制备糖肽。通过与肼的β-消除反应从蛋白质中释放三糖，并用2-氨基吡啶标记糖链的还原端。通过凝胶过滤和反相高效液相色谱法纯化三糖的荧光吡啶氨基衍生物。对所得到的糖肽和吡啶基氨基三糖进行了亚硫酸化， ...更多信息 进行甲基化研究。500-MHz ^1H核磁共振光谱和高碘酸盐氧化。通过在手性柱上的高效液相色谱法，葡萄糖和木糖属于D系列。根据结果，提出三糖的结构为：D-Xylpalpha 1 -3-D-Xylpalpha-1-3-D-Glcpbeta 1-O-Ser-53.2。蛋白Z是一种维生素K依赖性糖蛋白，从人和牛血浆中分离并表征。通过从肝cDNA文库中分离噬菌体克隆和体内扩增另外两个肝文库，获得了编码人蛋白Z的cDNA。蛋白质Z由40个氨基酸的前原前导序列合成。成熟蛋白由360个残基组成，包括13个羧基谷氨酸残基的Gla结构域、两个表皮生长因子结构域和与丝氨酸蛋白酶的催化结构域高度同源的羧基末端区。然而，人类蛋白Z在活性位点的催化三联体中含有Asp而不是Ser，含有Lys而不是His。少

项目成果

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”。

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。”

Yoshizumi,K.et al.: "Purification and Amino Acid Sequence of Basic Protein I,a Lysineー49ーPhospholipase A2 with Low Active,from the Venom of Trimeresurus flavoviridis (Habu Snake)," Toxicon. 28. 43-54 (1990)

Yoshizumi, K. 等人：“来自竹叶青（哈布蛇）毒液的碱性蛋白 I（一种低活性赖氨酸 49-磷脂酶 A2）的纯化和氨基酸序列”，Toxicon 28. 43-54（ 1990）

Tokunaga,F.et al.: "The NH_2ーterminal Residues of Rat Liver Proteosome (multicatalytic Proteinase complex) Subunits.C2,C3 and C8,are Naーacetylated." FEBS Lett.263. 373-375 (1990)

Tokunaga, F. 等人：“大鼠肝脏蛋白体（多催化蛋白酶复合物）亚基的 NH_2 末端残基。C2、C3 和 C8 被 Na 乙酰化。” FEBS Lett.263 (1990)。