The determination of amino acid sequence of human early pregnancy factor and the research on its bioactivity

人早孕因子氨基酸序列测定及其生物活性研究

• 批准号: 06671687
• 负责人: SUEOKA Kou
• 金额: $ 1.34万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671687/
• 关键词: EPF; Rosette inhibition test; SDS-PAGE; Glycoprotein; Western blot; RP-HPLC; Thioredoxin; Chaperonin 10

Human early pregnancy factor (EPF) has been elucidated in the biochemical characteristics, chemical structure and the bioactivity specially for the essential role of EPF in the mechanisms of establishment of pregnancy.The purified property from the source of human urine from pregnant women has been determined amino acid sequences from N-terminal of its peptide. Our purified substance with EPF bioactivity had 28KD of molecular weight and acidic peptide.The other purified properties of EPF are from the extract of human placenta had been recently reported by Clarke et al., which had the identified sequences with Thioredoxin (Adult type T cell leukemia factor ; ADF). And Chaperonin 10 homology peptide purified from human serum by Morton et al.which have 21KD of molecular weight and acidic peptide similar with our isolated peptide. In order to identify these rwo EPF properties, the polyclonal antibodies has been made by immunizind synthesized peptides with parts of EPF chemical structure to the rabbit. The response for biological activity (rosette inhibition activity) and serological responses (Dot blot, ELISA), with this antibody, and biochemical determination have demonstrated that these three EPF-bioactive properties were suggested to consists of completely different biochemical structures.

人类早孕因子（EPF）在生物化学特性、化学结构和生物活性等方面得到了阐明，特别是EPF在妊娠建立机制中的重要作用。从孕妇尿源纯化的性质已确定从其肽的n端氨基酸序列。纯化的EPF活性物质分子量为28KD，为酸性多肽。最近Clarke等人报道了从人胎盘提取物中纯化出的EPF的其他特性，其中鉴定出了与Thioredoxin （Adult type T cell leukemia factor； ADF）相关的序列。Morton等人从人血清中纯化的chaperonin10同源肽，分子量为21KD，酸性肽与我们分离的肽相似。为了鉴定这两种EPF的性质，我们将合成的EPF部分化学结构的肽段免疫兔制备了多克隆抗体。该抗体的生物活性（玫瑰抑制活性）和血清学反应（Dot blot， ELISA）以及生化测定表明，这三种epf生物活性特性可能由完全不同的生化结构组成。

项目成果

K.Sueoka, H.Asada, T.Hashiba, N.Kuji, T.Miyazaki, T.Kobayashi & S.Nozawa: "The falloposcopic findings of entire fallopian tube as a circumstance of fertilization and early development of embryos" journal of Fertilization and Implantation. 12 (1). 85-88 (1

K.Sueoka、H.Asada、T.Hashiba、N.Kuji、T.Miyazaki、T.Kobayashi

K.Sueoka: "Early pregnancy factor. (Basic and Clinical Finding.)" Nanzando, Tokyo, March 30. 207-214 (1995)

K.Sueoka：“早孕因素。（基本和临床发现。）”Nanzando，东京，3 月 30 日。207-214 (1995)

末岡 浩: "卵管-基礎から臨床まで-" 南山堂(in press), (1995)

Hiroshi Sueoka：“输卵管 - 从基础知识到临床实践”Nanzando（印刷中），（1995 年）

末岡浩: "受精および初期胚環境としての卵管内病態の検討-卵管鏡下卵管形成システムを用いて" 日本受精着床学会雑誌. 12. 85-88 (1995)

Hiroshi Sueoka：“作为受精和早期胚胎环境的输卵管内病理检查 - 使用输卵管镜输卵管形成系统”日本受精与植入学会杂志 12. 85-88 (1995)。

黒島正子: "Minimal damage embryo biopsyの技術的検討" 日本受精着床学会雑誌. 13(in press). (1996)

Masako Kuroshima：“微损伤胚胎活检技术研究”日本受精与植入学会杂志13（出版中）。