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-Structure paper
タイトル | Temperature-Resolved Cryo-EM Uncovers Structural Bases of Temperature-Dependent Enzyme Functions. |
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ジャーナル・号・ページ | J Am Chem Soc, Vol. 141, Issue 51, Page 19983-19987, Year 2019 |
掲載日 | 2019年12月26日 |
著者 | Chin-Yu Chen / Yuan-Chih Chang / Bo-Lin Lin / Chun-Hsiang Huang / Ming-Daw Tsai / |
PubMed 要旨 | Protein functions are temperature-dependent, but protein structures are usually solved at a single (often low) temperature because of limitations on the conditions of crystal growth or protein ...Protein functions are temperature-dependent, but protein structures are usually solved at a single (often low) temperature because of limitations on the conditions of crystal growth or protein vitrification. Here we demonstrate the feasibility of solving cryo-EM structures of proteins vitrified at high temperatures, solve 12 structures of an archaeal ketol-acid reductoisomerase (KARI) vitrified at 4-70 °C, and show that structures of both the Mg form (KARI:2Mg) and its ternary complex (KARI:2Mg:NADH:inhibitor) are temperature-dependent in correlation with the temperature dependence of enzyme activity. Furthermore, structural analyses led to dissection of the induced-fit mechanism into ligand-induced and temperature-induced effects and to capture of temperature-resolved intermediates of the temperature-induced conformational change. The results also suggest that it is preferable to solve cryo-EM structures of protein complexes at functional temperatures. These studies should greatly expand the landscapes of protein structure-function relationships and enhance the mechanistic analysis of enzymatic functions. |
リンク | J Am Chem Soc / PubMed:31829582 |
手法 | EM (単粒子) |
解像度 | 2.17 - 3.0 Å |
構造データ | EMDB-0740, PDB-6kou: EMDB-0742, PDB-6kpa: EMDB-0743, PDB-6kpe: EMDB-0746, PDB-6kph: EMDB-0747, PDB-6kpi: EMDB-0748, PDB-6kpj: EMDB-0749, PDB-6kpk: EMDB-0750, PDB-6kq4: EMDB-0751, PDB-6kq8: EMDB-0752, PDB-6kqj: |
化合物 | ChemComp-MG: ChemComp-NAI: ChemComp-9TY: |
由来 |
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キーワード | ISOMERASE / Complex |