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-Structure paper
タイトル | A 5+1 assemble-to-activate mechanism of the Lon proteolytic machine. |
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ジャーナル・号・ページ | Nat Commun, Vol. 14, Issue 1, Page 7340, Year 2023 |
掲載日 | 2023年11月13日 |
著者 | Shanshan Li / Kan-Yen Hsieh / Chiao-I Kuo / Tzu-Chi Lin / Szu-Hui Lee / Yi-Ru Chen / Chun-Hsiung Wang / Meng-Ru Ho / See-Yeun Ting / Kaiming Zhang / Chung-I Chang / |
PubMed 要旨 | Many AAA+ (ATPases associated with diverse cellular activities) proteins function as protein or DNA remodelers by threading the substrate through the central pore of their hexameric assemblies. In ...Many AAA+ (ATPases associated with diverse cellular activities) proteins function as protein or DNA remodelers by threading the substrate through the central pore of their hexameric assemblies. In this ATP-dependent translocating state, the substrate is gripped by the pore loops of the ATPase domains arranged in a universal right-handed spiral staircase organization. However, the process by which a AAA+ protein is activated to adopt this substrate-pore-loop arrangement remains unknown. We show here, using cryo-electron microscopy (cryo-EM), that the activation process of the Lon AAA+ protease may involve a pentameric assembly and a substrate-dependent incorporation of the sixth protomer to form the substrate-pore-loop contacts seen in the translocating state. Based on the structural results, we design truncated monomeric mutants that inhibit Lon activity by binding to the native pentamer and demonstrated that expressing these monomeric mutants in Escherichia coli cells containing functional Lon elicits specific phenotypes associated with lon deficiency, including the inhibition of persister cell formation. These findings uncover a substrate-dependent assembly process for the activation of a AAA+ protein and demonstrate a targeted approach to selectively inhibit its function within cells. |
リンク | Nat Commun / PubMed:37957149 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.4 - 5.8 Å |
構造データ | EMDB-34000, PDB-7yph: EMDB-34001, PDB-7ypi: EMDB-34002, PDB-7ypj: EMDB-34003, PDB-7ypk: EMDB-34004: Spiral hexamer of the substrate-free Lon protease with an S678A mutation EMDB-34005: Open-spiral pentamer of the substrate-free Lon protease with Y397A and S678A mutations EMDB-34006: Spiral hexamer of the substrate-free Lon protease with Y397A and S678A mutations EMDB-34107, PDB-7yuh: EMDB-34108, PDB-7yum: EMDB-34109, PDB-7yup: EMDB-34110, PDB-7yut: EMDB-34111, PDB-7yuu: EMDB-34112, PDB-7yuv: EMDB-34113, PDB-7yuw: EMDB-34114, PDB-7yux: EMDB-34116: Spiral pentamer of the substrate-free Lon protease with an M217A mutation EMDB-34117: Spiral hexamer of the substrate-free Lon protease with an M217A mutation EMDB-36865: Spiral pentameric form of the substrate-free Lon protease with a Y224S mutation in the presence of the N-terminal-truncated monomeric mutant bearing an E613K mutation EMDB-36866: Spiral hexameric form of the substrate-free Lon protease with a Y224S mutation in the presence of the N-terminal-truncated monomeric mutant bearing an E613K mutation EMDB-36867, PDB-8k3y: |
化合物 | ChemComp-AGS: ChemComp-ADP: |
由来 |
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キーワード | HYDROLASE / Lon protease / hydrolysis / AAA proteins / AAA / protease / complex / proteolysis / Assembly / AAA+ protein / ATPase |