Chemical biology study on protein phosphatases and their specific inhibitors.

蛋白磷酸酶及其特异性抑制剂的化学生物学研究。

• 批准号: 12045268
• 负责人: OSADA Hiroyuki
• 金额: $ 12.67万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12045268/
• 关键词: phoslactomycin; phosphatase; inhibitor; biosynthesis; apoptosis; dephosphorylation; isoavenaciolide; Bcl-2; リン酸化; 4-isoavenaciolide; VHR; Michael付加; 活性中心; RK-682; 分子設計; テトロン酸

Proteins involved in the signal transduction are regulated by phosphorylation/dephosphorylation during a process of proliferation, differentiation and apoptosis.Many selective inhibitors for protein kinases have been developed, however development of selective phosphatase inhibitors has not been achieved yet.To resolve this problem, I have tried to develop phosphatase inhibitors.1.4-isovavenaciolide was isolated as an inhibitor of the VHR protein tyrosine phosphatase. As 4-isoavenaciolide possesses a reactive exo-methylene moiety, it is possible that 4-isoavenaciolide inhibits VHR through the direct modification of a cysteine residue in the catalytic site.Since dual-specificity phosphatases have a consensus CXXXXXR motif in the catalytic site, dual-specificity phosphatases are sensitive to 4-isoavenaciolide.2.Phoslactomycins (PLMs) produced by Streptomyces, are potent and selective inhibitors of serine threonine phosphatases, PP2A.Incorporation studies with stable isotope labeling cyclohexanecarboxylic acid (CHC) indicated that the six PLM analoges (PLM A-F) made by Streptomyces sp.HK-803 are all biosynthesized from a CHC starter unit.Hydroxylation of the CHC-derived side chain of PLM B and a subsequent esterification produces the remaining PLM analogs.The entire 75 kb PLM biosynthetic gene cluster of the producing strain was cloned, sequenced and analyzed.Thecluster contains six genes (plml-7) which comprise the loading domain and seven extension modules of the type I modular PLM polyketide synthase.

参与信号转导的蛋白质在增殖、分化和凋亡过程中受到磷酸化/去磷酸化的调节。已经开发了许多选择性蛋白激酶抑制剂，但尚未开发出选择性磷酸酶抑制剂。为了解决这个问题，我尝试开发磷酸酶抑制剂。1.4-isoovavenaciolide作为一种 VHR 蛋白酪氨酸磷酸酶抑制剂。由于4-异燕麦内酯具有反应性外亚甲基部分，因此4-异燕麦内酯可能通过直接修饰催化位点中的半胱氨酸残基来抑制VHR。由于双特异性磷酸酶在催化位点中具有共有的CXXXXXR基序，因此双特异性磷酸酶对 4-isoavenaciolide.2. 由链霉菌产生的磷乳霉素 (PLM)，是丝氨酸苏氨酸磷酸酶 PP2A 的有效和选择性抑制剂。稳定同位素标记环己烷甲酸 (CHC) 的掺入研究表明，由链霉菌 sp.HK-803 产生的六种 PLM 类似物 (PLM A-F) 均是 由 CHC 起始单元生物合成。PLM B 的 CHC 衍生侧链的羟基化和随后的酯化产生剩余的 PLM 类似物。对生产菌株的整个 75 kb PLM 生物合成基因簇进行了克隆、测序和分析。该簇包含 6 个基因 (plml-7)，其中包含加载结构域和 7 个延伸域 I 型模块化 PLM 聚酮合酶的模块。

项目成果

Takanami-Ohnishi, Y. et al.: "Essential role of p38 mitogen-activated protein kinase in contact hypersensitivity."J.Biol.Chem.. 277. 37896-37903 (2002)

Takanami-Ohnishi, Y. 等人：“p38 丝裂原激活蛋白激酶在接触性超敏反应中的重要作用。”J.Biol.Chem.. 277. 37896-37903 (2002)

Cui, C.-B.et al.: "A new polyphenolic compound from Rubus aleaefolius and its inhibitory activity on mammalian cell cycle at GO/G1 phase."Chinese Chem.Lett.. 13. 327-330 (2002)

Cui, C.-B.et al.：“一种来自悬钩子的新多酚化合物及其对 GO/G1 期哺乳动物细胞周期的抑制活性。”Chinese Chem.Lett.. 13. 327-330 (2002)

Osada, H.: "An overview on the diversity of actinomycete metabolites."Actinomycetol.. 15. 11-14 (2001)

Osada, H.：“放线菌代谢物多样性概述。”Actinomycetol.. 15. 11-14 (2001)

Shoji, M., Kishida, S., Takeda, M., Kakeya, H., Osada, H., Hayashi, Y.: "A practical total synthesis of both enantiomers of epoxyquinols A and B."Tetrahedron Lett.. 43. 9155-1958 (2002)

Shoji, M.、Kishida, S.、Takeda, M.、Kakeya, H.、Osada, H.、Hayashi, Y.：“环氧醌醇 A 和 B 两种对映体的实用全合成。”Tetrahedron Lett.. 43

Nie, L., Ueki, M., Kakeya, H., Osada, H.: "A facile and effective screening method for p21 WAF1 promoter activators from microbial metabolites."J. Antibiot.. 54. 783-788 (2001)

Nie, L.、Ueki, M.、Kakeya, H.、Osada, H.：“一种从微生物代谢物中筛选 p21 WAF1 启动子激活剂的简便有效的方法。”