Study on development of drugs for endocrine and metabolic disorders based on properties of new enzyme

基于新酶特性的内分泌代谢疾病药物开发研究

• 批准号: 12672127
• 负责人: TAKAHASHI Noriko
• 金额: $ 1.73万
• 依托单位: Hoshi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672127/
• 关键词: Acetoacetyl-CoA synthetase; Ketone body; cDNA; Brain; Diabetes; Cholesterol; HMG-CoA reductase; Pravastatin; アセトアセチルCoA合成酵素; コレスチラミン; 肝臓; ヒト; ラット; アセト酢酸; クローニング

Acetoacetyl-CoA (AA-CoA) synthetase is a novel ligase, which specifically activates acetoacetate to AA-CoA. This enzyme is mainly involved in lipid and cholesterol biosynthesis in the liver, and the physiological role of AA-CoA synthetase is not clear yet. This let us to develop new drugs by the elucidation of new enzyme. First, cloning of the enzyme cDNA was carried out being based on the amino acid sequence determined from the enzyme preparation purified from rat liver. Consequently, both the obtained rat and human AA-CoA synthetase genes consisted of 2016 nucleotides, and coded 672 amino acid residues. Homology of amino acid sequence between rat and human was 89.3%. Expression of this enzyme was high in brain, and its expression profile was similar to that of HMG-CoA reductase, the rate-limiting enzyme of cholesterol biosynthesis. In human brain, expression of the enzyme was high in cerebral cortex, especially in hippocampus, which is located inner side part of temporal lobe, and expression profile of the AA-CoA synthetase was similar to that of HMG-CoA reductase. In diabetic rats injected with streptozotocin (STZ), hepatic AA-CoA synthetase activity started to gradually decrease similarly as HMG-CoA reductase and reached one tenth of the initial activity. When rats were fed with the diet supplemented with hypocholesterolemic agents, pravastatin and cholestyramine, the degree of STZ-induced decrease in AA-CoA synthetase activity was diminished, and ketone body concentration in blood was decreased to the normal level. These results indicate that hepatic AA-CoA synthetase may partly contribute to the regulation of blood ketone body, and that this enzyme expressed highly in hippocampus of human brain may have the important role in memory function.

乙酰乙酰辅酶A合成酶（Acetoacetyl-CoA synthetase，AA-CoA synthetase）是一种新型的连接酶，能特异性地激活乙酰乙酸生成AA-CoA。该酶主要参与肝脏中脂质和胆固醇的生物合成，AA-CoA合成酶的生理作用尚不清楚。这使我们能够通过对新酶的阐明来开发新药。首先，根据从大鼠肝脏纯化的酶制剂测定的氨基酸序列进行酶cDNA的克隆。因此，获得的大鼠和人AA-CoA合成酶基因均由2016个核苷酸组成，编码672个氨基酸残基。大鼠与人的氨基酸序列同源性为89.3%。该酶在脑中的表达较高，其表达谱与胆固醇生物合成的限速酶HMG-CoA还原酶的表达谱相似。在人脑中，该酶在大脑皮层，特别是位于颞叶内侧部分的海马中表达较高，并且AA-CoA合成酶的表达谱与HMG-CoA还原酶的表达谱相似。在注射链脲佐菌素（STZ）的糖尿病大鼠中，肝脏AA-CoA合成酶活性开始逐渐降低，类似于HMG-CoA还原酶，并达到初始活性的十分之一。当大鼠喂食补充有降胆固醇剂、普伐他汀和考来烯胺的饮食时，STZ诱导的AA-CoA合成酶活性降低的程度减轻，血液中的酮体浓度降低至正常水平。提示肝脏AA-CoA合成酶可能参与血酮体的调节，在人脑海马区高表达的该酶可能在记忆功能中起重要作用。

项目成果

Iwahori A., Takahashi N.et al.: "cDNA-derived amino acid sequence of acetoacetyl-CoA synthetase from rat liver"FEBS Letter. 466. 239-243 (2000)

Iwahori A.、Takahashi N.等人：“来自大鼠肝脏的乙酰乙酰辅酶A合成酶的cDNA衍生氨基酸序列”FEBS Letter。

Influence of Streptozotocin Diabetes on Acetoacetyl-CoA Synthetase in the Rat

链脲佐菌素对糖尿病大鼠乙酰乙酰辅酶A合成酶的影响

• 期刊: Biochemical Pharmacology (in press)
• 作者: Hiroki Sato;Noriko Takahashi;Mayumi Nakamoto;Masahiro Ohgami;Masahiro Yamasaki;Tetsuya Fukui
• 通讯作者: Tetsuya Fukui

cDNA-derived amino acid sequence of acetoacetyl-CoA synthetase from rat liver

大鼠肝脏乙酰乙酰辅酶A合成酶的cDNA衍生氨基酸序列

• 发表时间: 2000
• 期刊: FEBS Letter 466
• 作者: Iwahori A.;Takahashi;N;et al.
• 通讯作者: et al.

Influence of Streptozotocin Diabetes on Acetoacetyl-CoA, Synthetase in the Rat

链脲佐菌素对糖尿病大鼠乙酰乙酰辅酶A、合成酶的影响

• 期刊: Biochemical Pharmacology (in press)
• 作者: Sato;H.;Takahashi;N.;et al
• 通讯作者: et al

Sato, H., Takahashi, N.et al.: "Influence of Streptozotocin Diabetes on Acetoacetyl-CoA Synthetase in the Rat"Biochemical Pharmacology. (in press). (2002)

Sato, H.、Takahashi, N.等人：“链脲佐菌素糖尿病对大鼠乙酰乙酰辅酶A合成酶的影响”生化药理学。