Ornithine decarboxylase antizyme - Its structure, role, regulation, and comparative biochemistry

鸟氨酸脱羧酶抗酶 - 其结构、作用、调节和比较生物化学

• 批准号: 63480131
• 负责人: HAYASHI Shin-ichi
• 金额: $ 4.35万
• 依托单位: The Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480131/
• 关键词: Ornithine decarboxylase; Antizyme; cDNA; Translational regulation; Frame-shift; mRNA; Half-life; Polyamines; アンチザイムインヒビター; 酵素分解; ラット; カエル

Ornithine decarboxylase (ODC) antizyme is a unique regulatory protein involved in feedback inhibition and possibly degradation of the enzyme. Following results were obtained in this second year of supported research.(1) Determination of base sequence of antizyme MRNASequence analysis of newly cloned CDNA and genomic DNA indicated that rat liver antizyme is consisted of 227 amino acids and that translation of antizyme MRNA is likely to involve a frame-shift, a very unusual process in bukaryotic cells. (Matsufuji)(2) Tissue distribution of antizyme MRNANorthern blot analysis demonstrated that a single species of antizyme MRNA of 1.3 kb was widely and amply distributed in various murine tissues. The MRNA was quite stable in vivo (T_<1/2> = 12 h) and its amount did not increase upon putrescine treatment, indicating that the induction of antizyme by polyamines is due to translational stimulation. (Matsufuji)Role of antizymeProtein synthesis inhibitors like emetine and pactamycin inhibited antizyme synthesis as well as polyamine-caused acceleration of ODC decay, while amino acid analogs like ethionine and 5-fluorotryptophan inhibited neither antizyme induction nor polyamine-caused acceleration of ODC decay, indicating a close relationship between these two processes. An antizyme overproducing variant cell line was obtained from ODC-overproducing CHO cells. Studies on half-lives of free and antizyme-bound ODC in this cell line strongly suggested antizyme-mediated degradation of ODC. (Murakami)(4) Presence and role of antizyme in the frog Xenopus laevisInduction of antizyme and acceleration of ODC decay were both observed in primary cultured frog hepatocytes. (Hayashi & Murakami)

鸟氨酸脱羧酶（ODC）抗酶是一种独特的调节蛋白，参与反馈抑制和可能的酶降解。在第二年的资助研究中取得了以下成果。(1)抗酶MRNA的碱基序列测定及对新克隆的cDNA和基因组DNA的序列分析表明，大鼠肝抗酶由227个氨基酸组成，抗酶MRNA的翻译可能涉及到在细胞中非常罕见的移码过程。（2）抗酶的组织分布MRNANOR杂交分析表明，1.3kb的抗酶MRNA在小鼠的各种组织中分布广泛。该mRNA在体内相当稳定（T <12> = 12 h），经腐胺处理后其量不增加，表明多胺诱导抗酶是由于翻译刺激。抗酶的作用蛋白质合成抑制剂如embryonic和pactamycin抑制抗酶的合成以及多胺引起的ODC衰变的加速，而氨基酸类似物如etherylate和5-fluorotropophosphate既不抑制抗酶的诱导也不抑制多胺引起的ODC衰变的加速，表明这两个过程之间的密切关系。从过量产生ODC的CHO细胞获得过量产生抗酶的变体细胞系。对该细胞系中游离和抗酶结合的ODC的半衰期的研究强烈地表明了抗酶介导的ODC降解。（Murakami）（4）抗酶在非洲爪蟾肝细胞中的存在及其作用在原代培养的蛙肝细胞中，抗酶的诱导和ODC的加速衰变均被观察到。（Hayashi & Murakami）

项目成果

S. Matsufuji, R. Kanamoto, Y. Murakami and S. Hayashi: "Monoclonal antibody studies on the properties and regulation of murine ornithine decarboxylase antizymes." J. Biochem. 107: 87-91 (1990).

S. Matsufuji、R. Kanamoto、Y. Murakami 和 S. Hayashi：“关于鼠鸟氨酸脱羧酶抗酶特性和调节的单克隆抗体研究”。

S.Hayashi: "Ornithine Decarboxylase:Biology,Enzymology,and Molecular Genetics." Pergamon Press, 147 (1989)

S.Hayashi：“鸟氨酸脱羧酶：生物学、酶学和分子遗传学。”

S.Matsufuji: "Distribution and properties of ornithine decarboxylase antizyme.In S.H.Goldemberg and I.D.Algranati(eds.)The Biology and Chemistry of Polymines." IRL Press,Oxford, 81-90 (1989)

S.Matsufuji：“鸟氨酸脱羧酶抗酶的分布和特性。见 S.H.Goldemberg 和 I.D.Algranati（编辑）《多胺的生物学和化学》。”

S. Matsufuji, T.G. Baby, R. Kanamoto, T. Kameji, Y. Murakamiand S. Hayashi: "Distribution and properties of ornithine decarboxylase antizyme." In: The Biology and Chemistry of Polyamines. (S.H. Goldemberg and I.D. Algranati, eds.) IRL Press Oxford, pp. 81

S.松藤，T.G.

Y.Murakami: Biochem.J.254. 367-372 (1988)

Y.Murakami：Biochem.J.254。