Analysis of O-sulfotyrosine-mediated bio-interactions using synthetic peptides

使用合成肽分析 O-磺基酪氨酸介导的生物相互作用

基本信息

批准号：
13672241
负责人：
KITAGAWA Kouki
金额：
$ 2.3万
依托单位：
Nigata University of Phermacy and Applied Life Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

1 We carried out the chemical synthesis of large molecular forms of Tyr(SO_3H)-containing peptides based on the facile and efficient solid-phase method developed by us.i) Human big gastrin-II and its C-terminal Gly-extended peptide were prepared by the convergent segment condensation approach on the resin.ii) Human cholecystokinin (CCK)-58 was prepared by the silver-ion mediated thioester segment condensation approach. In this synthesis, the 2-chlorotrityl resin was extensively used as a solid support to prepare the sulfated segment and partially protected thioester segments. CCK-58 exhibited glucose-dependent insulinotropic activity at levels comparable to CCK-33.2 Three α-conotoxins (PnIA, PnIB, and EpI) that contain the sulfated tyrosine residue were synthesized by the Fmoc-based solid-phase method. Both a simultaneous oxidation approach and a two-step selective oxidation approach were employed to form the two disulfide linkages. In a simultaneous approach for PnIA and PnIB, additio … More n of DMSO in the reaction medium was critical to reduce the production of disulfide bond isomers. Desulfation was kept minimum throughout synthesis. α-Conotoxin EpI was synthesized using a novel solid-phase approach, in which α-carboxyl function of Asp was utilized as an anchoring group with a solid support. Three sulfated α-conotoxins were subjected to bioassay (Inhibitory effects of catecholeamine secretion from bovine adrenal chromaffin cells) and were found to be equipotent with their non-sulfate counterparts. This implies that the sulfate groups on these α-conotoxins are not determinants for their biological activities. Also we found that the disulfide bond isomers of these α-conotoxins had a weak but an apparent inhibitory effects of catecholeamine secretion.3 Various sized rat CCK-peptides (CCK-33, CCK-22, and CCK-12) were prepared by our solid-phase method and they were used as markers on the HPLC separations of the proteolytic products from proCCK. A model for the progression of pro-CCK processing in AtT 20-cells was proposed.4 Using the synthetic sulfated peptides, factor XI (FXI) was found to bind with the platelet glycoprotein Ibα not via an anionic duster in which three sulfated tyrosine residues were involved, but via the leucine-rich repeat sequences within the N-terminal domain of Gplbα. Less

1 基于我们开发的简便高效的固相方法，我们进行了大分子形式的含Tyr(SO_3H)肽的化学合成。 i) 人大胃泌素-II及其C端Gly延伸肽通过树脂上的会聚链段缩合方法制备。ii) 人胆囊收缩素 (CCK)-58 通过银离子介导的硫酯链段制备在该合成中，使用 2-氯三苯甲基树脂作为固体载体来制备硫酸化片段和部分受保护的硫酯片段，CCK-58 显示出与 CCK-33.2 三种 α-芋螺毒素大致相当的葡萄糖依赖性促胰岛素活性。含有硫酸化酪氨酸残基的（PnIA、PnIB和EpI）是通过基于Fmoc的固相方法同时合成的。采用氧化方法和两步选择性氧化方法形成两个二硫键。在 PnIA 和 PnIB 的同步方法中，反应介质中添加 DMSO 对于减少二硫键异构体的产生至关重要。使用新型固相方法合成α-芋螺毒素EpI，其中Asp的α-羧基功能被用作锚定基团对三种硫酸化 α-芋螺毒素进行了生物测定（牛肾上腺嗜铬细胞分泌儿茶酚胺的抑制作用），发现其与非硫酸盐立方体等效，这意味着这些 α-芋螺毒素上的硫酸基团是等效的。不是其生物活性的决定因素。我们还发现这些α-芋螺毒素的二硫键异构体具有较弱但明显的抑制作用。儿茶酚胺分泌。3 通过我们的固相方法制备了各种大小的大鼠 CCK 肽（CCK-33、CCK-22 和 CCK-12），并将它们用作 proCCK A 蛋白水解产物的 HPLC 分离的标记物。提出了 AtT 20 细胞中 pro-CCK 加工进程的模型。4 使用合成的硫酸化肽，发现因子 XI (FXI) 与血小板糖蛋白 Ibα 不是通过涉及三个硫酸化酪氨酸残基的阴离子分散体，而是通过 Gplbα Less 的 N 末端结构域内富含亮氨酸的重复序列。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Solid-phase synthesis of tyrosine sulfate containing α-conotoxins

含α-芋螺毒素硫酸酪氨酸的固相合成

DOI：
发表时间：
2003
期刊：
Peptides 2002
影响因子：
0
作者：
Yuushi Okumura;Kouki Kitagawa
通讯作者：
Kouki Kitagawa

Kouki Kitagawa: "Ionic interaction of the Tyr(SO_3H)residue with the cationic functional group in the biomolecule"Proceedings of the 4th International Symposium on Frontiers in Protein Chemistry and Biotechnology. 45-49 (2002)

北川幸树：“Tyr(SO_3H)残基与生物分子中阳离子官能团的离子相互作用”第四届国际蛋白质化学与生物技术前沿研讨会论文集。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Kinetic analysis of the interaction between vitronectin and the urokinase receptor

DOI：
10.1074/jbc.m111225200
发表时间：
2002-03-15
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Okumura, Y;Kamikubo, Y;Loskutoff, DJ
通讯作者：
Loskutoff, DJ

Synthesis of Tyr(SO_3H)-containing α-conotoxins

含Tyr(SO_3H)的α-芋螺毒素的合成

DOI：
发表时间：
2003
期刊：
Peptide Science 2002
影响因子：
0
作者：
Yuushi Okumura;Kouki Kitagawa;Yumi Sekigawa;Yuushi Okumura et al.;Kouki Kitagawa et al.;Yumi Sekigawa et al.
通讯作者：
Yumi Sekigawa et al.

Ionic interaction of the Tyr(SO3H) residue with the cationic Functional group in the biomolecule

Tyr(SO3H) 残基与生物分子中阳离子官能团的离子相互作用

DOI：
发表时间：
2002
期刊：
Frontiers in Protein Chemistry and Biotechnology
影响因子：
0
作者：
Yuushi Okumura;Kouki Kitagawa;Yumi Sekigawa;Yuushi Okumura et al.;Kouki Kitagawa et al.;Yumi Sekigawa et al.;Kouki Kitagawa
通讯作者：
Kouki Kitagawa