Research for structure-function relationship in human gonadotropins

人类促性腺激素结构与功能关系的研究

• 批准号: 02454379
• 负责人: SUGANUMA Nobuhiko
• 金额: $ 4.42万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454379/
• 关键词: human chorionic gonadotropin; luteinizing hormone; follicle stimulating hormone; beta subunit; disulfide bond; site-directed mutagenesis; expression vector; gene expression; ヒト繊毛性ゴナドトロピン; 免疫学的活性; 生物学的活性; SーS結合; 発現ベクタ-; ヒト絨毛性コナドトロピン

To analyze the intracellular roles of disulfide bonds in human gonadotropins, the cystein residues in each beta subunit were converted to other amino acid sequences by site-directed mutagenesis. The following results were obtained using the mutant beta subunits.1. Assembly with alpha subunit was blocked by the elimination of any disulfide bonds in hCG beta subunit, however, there were differences in the binding rate among each disulfide pair.2. The disulfide bond at cysteins 26/110 had regulating role in secretions of beta subunits of hCG and LH,but not in FSH beta subunit.3. Various immunological reactivities were observed when the disulfide bond at 23/72 in hCGbeta were eliminated.4. Receptor bindings and biological activities were not affected by disulfide bond breakage in hCGbeta.5. The process of disulfide bond formation in hCGbeta were also determined. These methods presented here could be useful to analyze the relationship between structure and function even in other peptides.

为了分析二硫键在人促性腺激素中的细胞内作用，每个β亚基中的半胱氨酸残基通过位置定向的诱变转化为其他氨基酸序列。使用突变β亚基获得以下结果。1。通过消除HCGβ亚基中的任何二硫键阻断，用α亚基组装被阻断，但是，每种二硫键对之间的结合速率差异。2。 Cysteins的二硫键26/110在HCG和LH的β亚基分泌中具有调节作用，但在FSHβ亚基中没有调节。3。当消除HCGBETA中23/72的二硫键键时，观察到了各种免疫反应率。4。 HCGBETA中的二硫键断裂不影响受体结合和生物学活性。5。还确定了HCGBETA中二硫键形成的过程。此处介绍的这些方法即使在其他肽中也可以分析结构和功能之间的关系。

项目成果

Nobuhiko Suganuma: "Intracellular roles of disulfide bonds in human chorionic gonadotropin β subunitーAnalysis using siteーdirected mutagenesis and gene transfer" 2nd Lake Shirakaba Placenta Conference. 39-53 (1991)

Nobuhiko Suganuma：“人绒毛膜促性腺激素 β 亚基中二硫键的细胞内作用 - 使用定点诱变和基因转移进行分析”第二届白桦湖胎盘会议。

Bedows E,Huth JR,Suganuma N,Bartels Cf,Boime I,Ruddon RW: "Disulfide bond mutations affect the folding of the human chorionic gonadotropin (hCG)-beta subunit in transfected CHO cells." J Biol Chem. 1993

Bedows E、Huth JR、Suganuma N、Bartels Cf、Boime I、Ruddon RW：“二硫键突变影响转染 CHO 细胞中人绒毛膜促性腺激素 (hCG)-β 亚基的折叠。”

Shiono S,Suganuma N,Bo M,Boime I,Seibert K,Nakao K.Mukoyama M,Imura H,Needleman P: "Post-translational processing and secretory pathway of human atriopeptin in rat pheochromocytoma cells." Bicohem Biophys Res Commun. 176. 1232-1238 (1991)

Shiono S，Suganuma N，Bo M，Boime I，Seibert K，Nakao K.Mukoyama M，Imura H，Needleman P：“大鼠嗜铬细胞瘤细胞中人心房肽的翻译后加工和分泌途径。”

Nobuhiko Suganuma: "Intracellular roles of disulfide bonds in human chorionic gonadotropin β subuniz ーAnalysis using siteーdirected mutagenesis and gene transferー" Lake Shirakaba Placenta Conference. 2. (1991)

Nobuhiko Suganuma：“人绒毛膜促性腺激素 β 亚基中二硫键的细胞内作用 - 使用定点诱变和基因转移进行分析 -”白桦湖胎盘会议 2。（1991）

F.A.Fares: "Design of a long-acting follitropin agonist by fusing the C-terminal sequence of the chorionic gonadocropin β subunit to the follitropin β subunit." Proc.Natl.Acad.Soi.U.S.A. 89. 4304-4308 (1992)

F.A.Fares：“通过将绒毛膜促性腺激素 β 亚基的 C 端序列与促卵泡素 β 亚基融合来设计长效促卵泡素激动剂。”Proc.Natl.Acad.Soi.U.S.A. 89. 4304-4308 (1992)