NEW FLUOROGENIC SYNTHETIC SUBSTRATES AND INHIBITORS FOR MICROSOMAL METALLOENDOPEPTIDASE

微粒体金属内肽酶的新型荧光合成底物和抑制剂

• 批准号: 04558025
• 负责人: KAWABATA Shun-ichiro
• 金额: $ 3.46万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04558025/
• 关键词: Processing protease; Metalloendopeptidase; Synthetic substrate; Fluorogenic substrate; Metalloprotease; プロペプチド酵素; プロセシング; メタルプロテアーゼ; 蛍光性ペプチド基質; 合成阻害剤

Ten fluorogenic synthetic substrates of Abz (2-aminobenzoyl)-peptide-Dna(2,4-dinitro-anilinoethylamide) were newly synthesized and tested to find specific substrates for rabbit liver microsomal metalloendopeptidase (MEP) as shown in Table. Among these substrates, Abz-ARVRRANS-Dna or Abz-ARVTRANS-Dna was found to be the most sensitive substrates for MEP.The assay method using these peptide substrates enabled us to detect MEP activity in an aliquot (20-50 mul) of the crude extract.

新合成了10种ABZ(2-aminobenzoyl)-peptide-Dna(2，4-dinitro-anilinoethylamide)的荧光合成底物，并对其进行了检测，以寻找兔肝微粒体金属内肽酶的特异性底物，如表所示。在这些底物中，ABZ-ARVRRANS-DNA或ABZ-ARVTRANS-DNA是对MEP最敏感的底物。使用这些底物的测定方法使我们能够在等量(20-50微升)的粗提液中检测到MEP活性。

项目成果

Ikura, T., Go, N., Kohda, D., Inagaki, F., Yanagawa, H., Kawabata, M., Kawabata, S., Iwanaga, S., Noguti, T., and Go. M.: "Secondary Structural Features of Modules M2 and M3 of Barnase in Solution by NMR Experiment and Distance Geometry Calculation" Prote

Ikura, T.、Go, N.、Kohda, D.、Inagaki, F.、Yanakawa, H.、Kawabata, M.、Kawabata, S.、Iwanaga, S.、Noguti, T. 和 Go。

Miura, Y., Kawabata, S., and Iwanaga, S.: "A Limulus Intracelluar Coagulation Inhibitor with Characteristics of the Serpin Superfamily, Purification, Characterization, and cDNA Cloning." J.Biol.Chem.269. 542-547 (1994)

Miura, Y.、Kawabata, S. 和 Iwanaga, S.：“具有丝氨酸蛋白酶抑制剂超家族特征的鲎细胞内凝血抑制剂、纯化、表征和 cDNA 克隆。”

Ikura,T.: "Secondary Structural Features of Modules M2 and M3 of Barnase in Solution by NMR Experiment and Distance Geometry Calculation." Proteins. 16. 341-356 (1993)

Ikura,T.：“通过 NMR 实验和距离几何计算研究溶液中 Barnase 模块 M2 和 M3 的二级结构特征。”

Sakumi,K.: "Cloning and Expression of cDNA for a Human Enzyme That Hydrolyzes 8-Oxo-dGTP,a Mutagenic Substrate for DNA Synthesis." The Journal of Biological Chemistry. 268. 23524-23530 (1993)

Sakumi,K.：“水解 8-Oxo-dGTP（一种用于 DNA 合成的诱变底物）的人类酶 cDNA 的克隆和表达。”

Sakumi, K., Furuichi, M., Tsuzuki, T., Kakuma, T., Kawabata, S., Maki, H., and Sekiguchi, M.: "Cloning and Expression of cDNA for a Human Enzyme That Hydrolyzes 8-Oxo-dGTP, a Mutagenic Substrate for DNA Synthesis." J.Biol.Chem.268. 23524-23530 (1993)

Sakumi, K.、Furuichi, M.、Tsuzuki, T.、Kakuma, T.、Kawabata, S.、Maki, H. 和 Sekiguchi, M.：“水解 8- 的人类酶的 cDNA 的克隆和表达