牛肝臓20Sプロテアソームの2.5Å分解能結晶構造解析

牛肝20S蛋白酶体2.5Å分辨率晶体结构分析

• 批准号: 13033034
• 负责人: 森本 幸生
• 金额: $ 2.05万
• 依托单位: Himeji Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13033034/
• 关键词: プロテアソーム; 超分子複合体; X線結晶構造解析; 免疫応答; 核内移行シグナル

獲得免疫を持った高等動物のプロテアソームは28個のサブユニットから成る多機能蛋白質分解酵素複合体であり、単に不要蛋白質を分解するだけではなく、内在性抗原を正確に選別し、抗原ペブチドを生成しで抗原提示を担う免疫応答系超分子システムである。本研究では我々がSPring-8放射光実験で得た牛肝臓20Sプロテアソームの2.5Å分解能でのX線データによる解析、精密化を通した超分子複合体としての蛋白分解機構と、抗原提示のための蛋白選別機構を能力を、その2.5Å分解能の結晶構造解析にもとづいて立体構造から考察する。また、これと比較して脾臓から抽出した免疫型20Sプロテアソームの構造解析にも着手する。牛肝臓から抽出したプロテアソームをMPDを沈殿剤として結晶化を行い、SPring-8大型放射光施設で回折実験を行った。以前得られていた結晶(Y.Morimoto, J.B.(1995))とは異なる晶系の結晶が得られ、同じ精製方法、結晶化であるにも関わらず異なる晶系、格子長の変化は、プロテアソームの分子状態が異なるものと考えられた(Y.Tomisugi, J.B.(2000))。SPring-8にて2.5Å分解能までの質の良い回折データを得ることができ、モデルを構築した。高等動物のプロテアソームに特有と見られる粒子内の塩基性アミノ酸残基の分布などが見られた。また構成型プロテアソームと免疫型プロテアソームのサブユニットの違い、あるいは保存されているサブユニットと活性機構の関連などを明らかにし考察した(M.Unno, J.B.(2002))。これとともにプロテアソームが細胞核内に移行されるときのシグナル(NLS)の分子表面および分子中での存在箇所を明らかにし、核内移行に対する疑問点を考察した(M. Unno, Structure,(2002))。

It was obtained that the immunized mice were able to carry out the production of multi-functional proteolytic enzyme complexes, proteolytic enzymes, internal antigens, and antigen production strategies in 28 immunized mice. In this study, we obtained the results of SPring-8 radioluminescence that the decomposition of bovine liver 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s, 20s The spleen was extracted from the spleen and was extracted from the spleen. The immune type 20s was extracted from the spleen. The extraction of the bovine liver, the extraction of the MPD, the crystallization of the crystal, the large-scale radioluminescence of SPring-8, the back-folding of the radiation equipment. (Y.Morimoto, J.B. (1995)). The results show that the crystal system of the crystal system is characterized by the method of crystallization, the method of crystallization, the crystallization of the crystal system, the lattice growth, the molecular morphology and the molecular structure of the crystal system (Y.Tomisugi, J.B. (2000)). SPring-8 two and a half can be decomposed to improve the quality of the product. The distribution of acid residues in particles is unique to the distribution of acid residues in particles. The immunoassay was used to detect the activity of the active mechanism (M.Unno, J.B. (2002)). Intranuclear migration (NLS), intranuclear migration (Unno, Structure, (2002)).

项目成果

K.Suto, et.al.: "High Resolution Structure of FMN-binding Protein"J.Phys.Soc.Jpn.. 70. 406-407 (2001)

K.Suto等：“FMN结合蛋白的高分辨率结构”J.Phys.Soc.Jpn..70.406-407(2001)

M.Unno, et al.: "The Structure of the Mammalian 20S Proteasome at 2.75Å Resolution"Structure. 10. 609-618 (2002)

M.Unno 等人：“2.75Å 分辨率下的哺乳动物 20S 蛋白酶体的结构”结构。10. 609-618 (2002)

M.Yamanishi, et al.: "The crystal structure of coenzyme B12-dependent glycerol dehydratse in complex with cobalamin and 1,2-propanediol"European J.Biochemistry. (in press). (2002)

M.Yamanishi 等人：“辅酶 B12 依赖性甘油脱水酶与钴胺素和 1,2-丙二醇复合物的晶体结构”European J.Biochemistry。

M.Unno, et.al.: "Structure determination of the Constitutive 20S proteasome from Bovine Liver at 2.75Å Resolution"J.Biochem.. 131. 171-173 (2002)

M.Unno 等人：“以 2.75Å 分辨率测定牛肝组成型 20S 蛋白酶体的结构”J.Biochem.. 131. 171-173 (2002)

Y.Tomisugi, et.al.: "New crystal forms and low resolution structure analysis of 20S Proteasomes from bovine liver"J.Biochem.. 127. 941-943 (2000)

Y.Tomisugi 等人：“来自牛肝的 20S 蛋白酶体的新晶体形式和低分辨率结构分析”J.Biochem.. 127. 941-943 (2000)