Research on the structure and function of antihemorrhagic factor

抗出血因子的结构与功能研究

• 批准号: 05660103
• 负责人: YONAHA Kazuo
• 金额: $ 1.34万
• 依托单位: University of The Ryukyus
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 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-05660103/
• 关键词: snake venom; hemorrhagic; antihemorrhagic; protein; evolution; structure; mongoose; mRNA; cDNA; 一次構造

This research has been taken by aiming at the development of a novel antitoxin with antihemorrhagic factor of mongoose. Three antihemorrhagic factors of mongoose were purified and characterized in this research. The three factors, AHF1, AHF2, and AHF3, are glycoproteins of monomer structure. AHF1 completely inhibited the hemorrhagic activity of HR2b, hemorrahgic factor of Habu snake. The antihemorrhagic factor formed a complex with the hemorrahgic facor. No alteration in the primary or secondary structures of both factors was demonstrated before and after incubation of both factors.The three antihemorrhagic factors are composed of about 600 amino acids of similar composition. AHF1 and AHF2 were of the same amino terminal amino acid sequence, but AHF3 showed different sequence, though high homology was found between AHF3 and the other two antihemorrhagic factors. The amino acid sequence of AHF1 covering 75.9% of whole sequence of human alpha1B-glycoprotein (alpha1B-G) showed 46.1% identity with the sequece of alpha1BG.These results suggest for the antihemorrhagic factor to be a homologous protein with human alpha1B-G and therefore a supergene family of immunoglobulin, and the different ancestral proteins for the antihemorrhagic factors of mammals and repitiles.The total RNA was successfully prepared from the liver of mongoose, and mRNA was isolated from the RNA preparation. The cDNA library containing 10^<10>/ml independent phage particle was constructed.

这项研究是通过瞄准了伴有抗头性因子的新型抗毒素的开发来进行的。在这项研究中纯化并表征了猫鼬的三种抗血症因子。 AHF1，AHF2和AHF3的三个因素是单体结构的糖蛋白。 AHF1完全抑制了HR2B（HABU蛇出血因子）的出血活性。抗疾病因子与出血面形成了复合物。在这两个因素孵育之前和之后，两种因素的主要或次级结构均未改变。这三个抗毒素因子由约600个相似组成的氨基酸组成。 AHF1和AHF2具有相同的氨基末端氨基酸序列，但AHF3显示出不同的序列，尽管在AHF3和其他两个抗Hagragic因子之间发现了高同源性。 AHF1的氨基酸序列覆盖了人α1b-糖蛋白（α1b-g）的整个序列的75.9％，与α1bg的序列显示出46.1％的认同。这些结果表明，这些结果表明，这些对抗毒素的蛋白质与超级蛋白质的同源性蛋白质有关哺乳动物和重点的抗血肿因子。总RNA从猫豆的肝脏中成功制备，并从RNA制备中分离出mRNA。构建了包含10^<10>/ml独立噬菌体粒子的cDNA库。

项目成果

Zhi-Qing Qi, Kazuo Yonaha, Yasuhiro Tomihara and Seizen Toyama: "Characterization of the antihemorrhagic factorof mongoose (Herpestes edwardsii)" Toxicon. 32 (11). 1459-1469 (1994)

Zi-Qing Qi、Kazuo Yonaha、Yasuhiro Tomihara 和 Seizen Toyama：“猫鼬（Herpestes edwardsii）抗出血因子的特征”Toxicon。

Zhi-Aing Qi: "Characterization of the antihemorrhagic factor of mongoose(Herpestes edwordsii)" Toxicon. 32. 1459-1469 (1994)

齐志爱：“猫鼬（Herpestes edwordsii）抗出血因子的表征”Toxicon。

Zhi-Qing Qi: "Isolation of homologous peptides to domains of human α1B-glycoprotein from a mongoose antihemorrhagic factor" Toxicon. 33(印刷中). (1995)

齐志清：“从猫鼬抗出血因子中分离人 α1B 糖蛋白结构域的同源肽”Toxicon 33（出版中）。

Zhi-Aing Qi: "Isolation of peptides homelogeus to domains of human HB-glycoprotein from a mongoose antihemonhogic factor" Toxicon. 33. 241-245 (1995)

齐志爱：“从猫鼬抗血因子中分离出与人 HB 糖蛋白结构域同源的肽”Toxicon。

Zhi-Qing Qi, Kazuo Yonaha, Yasuhiro Tomihara and Seizen Toyama: "Isolation of peptides homologous to domains of human alpha1B-glycoprotein from a mongoose antihemorrhagioc factor" Toxicon. 33 (2). 241-245 (1995)

Zi-Qing Qi、Kazuo Yonaha、Yasuhiro Tomihara 和 Seizen Toyama：“从猫鼬抗出血因子中分离与人 α1B-糖蛋白结构域同源的肽”Toxicon。