Bilogical Roles on N-Acylamino Acid Releasing-enzyme-especially on the protein myristylation-

N-酰基氨基酸释放酶的生物学作用-特别是对蛋白质肉豆蔻酰化-

• 批准号: 61580144
• 负责人: TSUNASAWA Susumu
• 金额: $ 1.34万
• 依托单位: Osaka 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: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61580144/
• 关键词: Acylamino acid releasing-enzyme; ミリスチル化

To elucidate the biological roles on N-acylamino acid releasing-enzyme, its substrate specificity was at first studied with acetylmethionyl amino acids, acetylalanyl peptides ranging from dithrough penta-alanine, and N-acylmethionylalanine having various acyl groups, and the following results were thus found. 1) The enzyme hydrolyses all the peptides tested except for acetyl-L-methionuyl-L-proline, -L-aspartic acid and -D-amino acids. 2) Among acetylated derivatives of oligoalanine, acetyl di- and trialanine were most rapidly hydrolyzed by the enzyme. 3) The enzyme removes more effectively acetyl or propionyl amino acid from various N-acylated peptides. As the second step, the determination of the primary structure of the enzyme was tried. Amino acid sequences of several peptides generated from the enzyme from pig liver, by digestion with Achromobacter protease I were partially determined, and the isolation of cDNA encoding the enzyme has been now performing in the cDNA library constructed from pig liver using a 24-mer nucleotide corresponding to the sequence partially determined as above, Glu-Pro-Glu-Glu-Ala-Ala-Ala-Leu-Tyr, as a prove.

为了阐明N-酰基氨基酸释放酶的生物学作用，首先用乙酰甲硫氨酰氨基酸、从二至五丙氨酸的乙酰丙氨酰肽以及具有各种酰基的N-酰基甲硫氨酰丙氨酸研究了其底物特异性，并由此发现了以下结果。 1) 该酶可水解所有测试的肽，但乙酰基-L-甲硫氨酰-L-脯氨酸、-L-天冬氨酸和-D-氨基酸除外。 2) 在低聚丙氨酸的乙酰化衍生物中，乙酰二丙氨酸和三丙氨酸被酶水解得最快。 3) 该酶更有效地去除各种N-酰化肽中的乙酰基或丙酰基氨基酸。第二步，尝试测定酶的一级结构。通过用无色杆菌蛋白酶 I 消化，由来自猪肝的酶产生的几种肽的氨基酸序列被部分确定，并且编码该酶的 cDNA 的分离现在已经在由猪肝构建的 cDNA 文库中进行，使用对应于如上所述部分确定的序列的 24 聚体核苷酸， Glu-Pro-Glu-Glu-Ala-Ala-Ala-Leu-Tyr，作为证明。

项目成果

Sato,Takaaki: Gene. 50. 247-257 (1986)

佐藤贵明：吉恩。

TSUNASAWA, Susumu: Tanpakushitsu Kogaku Nyumon <section>5. Sequence Analysis of the Protein roduced by Gene Eenginering). Syujunsha, 136-144 (1987)

纲泽进：Tanpakushitsu Kogaku Nyumon <部分>5。

Ikeuchi,Toshihiko: Eur.J.Biochem.167. 233-238 (1988)

池内俊彦：Eur.J.Biochem.167。

SATAO, Takaaki: "Expression of the Human Salivary <alpha>-Amylase Gene in Yeast and Characterization of the Secreted Protein" Gene. 50. 247-257 (1986)

SATAO，Takaaki：“人类唾液<α>-淀粉酶基因在酵母中的表达和分泌蛋白的表征”基因。

IKEUCHI, Toshihiko: "Purification and Characterizatin of the spoOA Protein of Bacillus substitlis from an Overproducing Strain of Escherichia coli" Eur. J. Biochem.167. 233-238 (1988)

池内俊彦：“来自过量生产的大肠杆菌菌株中枯草芽孢杆菌 spoOA 蛋白的纯化和表征”Eur。