Structure and Cell Biology of a Novel Multicatalytic Proteinase Complex

新型多催化蛋白酶复合物的结构和细胞生物学

• 批准号: 01304029
• 负责人: ICHIHARA Akira
• 金额: $ 7.04万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Co-operative Research (A)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01304029/
• 关键词: Proteasomes; Multicatalytic protease; Structural analysis; Multisubunit complex; High molecular weight protease; 分子遺伝学; 多機能プロテア-ゼ複合体; 細胞内プロテア-ゼ; 20S環状粒子; cDNAクロ-ニング; 発生と分化

Multicatalytic proteinase complex (Proteasomes) is a large assembly of many protein components with various protease activities. In 1990 we carried out a co-operative research to investigate structure and functions of proteasomes.1. Structural analysis --- cDNAs of 15 components of the proteasomes from yeast, rat and human were cloned and sequenced. These sequences showed no homology with any known proteins indicating that the proteasomes consist of subunits which are new protein family. This consideration is supported by the facts that homology among subunits from different animals was very high. Assembly of subunits to form a ring-shape particle can be observed by electron microscopy and similarity of shape among proteasomes of different animals suggests that structure of complex is important for the function.2. Functions of proteasomes --- It was shown that proteasomes play important role in embryogenesis, particularly maturation and fertilization of egg. The expression of proteasome mRNA was very high in cancerous tissues of liver, kidney and blood cells. It was also found that proteasomes translocate etween cytosol and nucleus during cell growth. These results indicate that proteasomes relate closely with cell growth and differentiation. This notion was supported by experiments of gene disruption of yeast subunits. Yeast received disrupted genes became lethal.

多催化蛋白酶复合物（蛋白酶体）是许多具有各种蛋白酶活性的蛋白质成分的大集合体。 1990年我们开展了一项合作研究，研究蛋白酶体的结构和功能。 1．结构分析——对来自酵母、大鼠和人类的蛋白酶体的 15 种成分的 cDNA 进行了克隆和测序。这些序列与任何已知蛋白质没有同源性，表明蛋白酶体由新蛋白质家族的亚基组成。来自不同动物的亚基之间的同源性非常高的事实支持了这种考虑。通过电子显微镜可以观察到亚基组装形成环形颗粒，不同动物蛋白酶体形状的相似性表明复合物的结构对其功能很重要。2．蛋白酶体的功能——研究表明，蛋白酶体在胚胎发生，特别是卵子的成熟和受精过程中发挥着重要作用。肝、肾、血细胞等癌组织中蛋白酶体mRNA表达量非常高。还发现细胞生长过程中蛋白酶体在细胞质和细胞核之间易位。这些结果表明蛋白酶体与细胞生长和分化密切相关。这一观点得到了酵母亚基基因破坏实验的支持。受到破坏基因的酵母变得致命。

项目成果

Atsushi Kumatori: "Abnormally high expression of proteasomes in human leukemic cells" Proc.Natl.Acad.Sci.U.S.A.87. 7071-7075 (1990)

Atsushi Kumatori：“人类白血病细胞中蛋白酶体的异常高表达”Proc.Natl.Acad.Sci.U.S.A.87。

K.Tanaka: "Direct Evidence for Nuclear and Cytoplasmic Colocation of Proteasomes(Multiprotease Complexes)in Liver." J.Cell.Physiol.139. 34-41 (1989)

K.Tanaka：“肝脏中蛋白酶体（多蛋白酶复合物）核和细胞质共定位的直接证据。”

Y.Saitoh: "Sodium dodecyl sulfateーinduced conformational and enzymatic changes of multiーcatalytic proteinase" Biochem.Biophys.Res.Communi.162. 334-339 (1990)

Y.Saitoh：“十二烷基硫酸钠诱导的多催化蛋白酶的构象和酶促变化”Biochem.Biophys.Res.Communi.162 (1990)。

Tamura, T., Lee, D. H., Osaka, F., Fujiwara, F., Shin, S., Chung, C. H., Tanaka, T. and Ichihara, A.: "Molecular cloning and sequence analysis of cDNAs for five major subunits of human liver proteasomes (multi-catalytyic proteinase complexes)." Biochim. B

Tamura, T.、Lee, D. H.、Osaka, F.、Fujiwara, F.、Shin, S.、Chung, C. H.、Tanaka, T. 和 Ichihara, A.：“五个主要亚基 cDNA 的分子克隆和序列分析

Tomohiro Tamura: "Molecular cloning and sequence snalysis of cDNAs for five major subunits of human liver proteasomes(multiーcatalytyic proteinase complexes)" Biochim.Biophys.Acta. (1991)

Tomohiro Tamura：“人肝蛋白酶体（多催化蛋白酶复合物）五个主要亚基的 cDNA 的分子克隆和序列分析”Biochim.Biophys.Acta（1991）。