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A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes Vol 437|27 October 2005|doi:10.1038/nature04106 Yuko Hirano 生科四甲 91390355.

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Presentation on theme: "A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes Vol 437|27 October 2005|doi:10.1038/nature04106 Yuko Hirano 生科四甲 91390355."— Presentation transcript:

1 A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes Vol 437|27 October 2005|doi:10.1038/nature04106 Yuko Hirano 生科四甲 91390355 葉孟軒

2 Model for the Structure of the 26S Proteasome

3 名詞解釋 26S Proteasome : 真核生物體中一種蛋白質水解酶,為 multisubunt ,是由 20S 的核心 proteasome 和兩軸上的 19S regulatory complexs 組合而成,而 20S 是由 28 個 subunits 組 成, ”α 1-7 β 1-7 β 1-7 α 1-7 ” 為其排列成圓筒狀方式 PAC1 & PAC2 : Proteasome assembling chaperone-1 & 2 ,為此 篇論文討論的重點, PAC1 和 PAC2 會形成一個 heterodimers ,能幫 助 proteasome precursors 聚集成為成熟的 preteasome MG132 :

4 實驗方法 DNA constructs and cell culture. Protein extracts, immunological analysis and antibodies. Glycerol gradient analysis Purification of PAC1-PAC2 complex Pulse-chase experiments RNAi experiments Assay of proteasome activity Chromatography

5 density

6 Introduction “ Half-proteasome ” 包括一個 α ring + 一個 β ring + heterodimeric 還有一個 chaperone Ump1 。要組成一個 成熟的 proteasome ,就需要兩個 Half-proteasome dimerize ,在結合之前 β ring 上的 propeptide 會先脫離, 而且 Ump1( 人體中為 hUmp1) 也會降解離去;在整個 assembling 過程當中, heterodimeric 所扮演的角色就 是促進 precursor proteasome 構型組成,此外還能提供 一個鷹架,協助 α ring 組成,還有保持充足的 α rings 以構成 half-proteasome ,接下來就是要利用實驗的方 法去證實 heterodimeric 的這些功能。

7 圖一 PAC-1 and PAC-2 associate with precursor proteasomes. 4~24%

8 圖二 PAC-1 and PAC-2 associate with precursor proteasomes.

9 The PAC1–PAC2 heterodimer is rapidly degraded by proteasomes. 圖三 4~17%

10 The PAC1–PAC2 heterodimer is rapidly degraded by proteasomes. 圖四

11 siRNA-mediated knockdown of PAC1 and PAC2 impairs proteasome assembly. 4~24%

12 圖六 PAC1–PAC2 provides a scaffold for a-ring formation.

13 圖六 PAC1–PAC2 provides a scaffold for a-ring formation.


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