6KPH
343 K cryoEM structure of Sso-KARI in complex with Mg2+, NADH and CPD
Summary for 6KPH
| Entry DOI | 10.2210/pdb6kph/pdb |
| Related | 6KOU |
| EMDB information | 0746 |
| Descriptor | Ketol-acid reductoisomerase, MAGNESIUM ION, 1,4-DIHYDRONICOTINAMIDE ADENINE DINUCLEOTIDE, ... (4 entities in total) |
| Functional Keywords | complex, isomerase |
| Biological source | Saccharolobus solfataricus |
| Total number of polymer chains | 12 |
| Total formula weight | 456888.06 |
| Authors | Chen, C.Y.,Chang, Y.C.,Lin, B.L.,Huang, C.H.,Tsai, M.D. (deposition date: 2019-08-15, release date: 2020-03-25, Last modification date: 2024-03-27) |
| Primary citation | Chen, C.Y.,Chang, Y.C.,Lin, B.L.,Huang, C.H.,Tsai, M.D. Temperature-Resolved Cryo-EM Uncovers Structural Bases of Temperature-Dependent Enzyme Functions. J.Am.Chem.Soc., 141:19983-19987, 2019 Cited by PubMed Abstract: 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. PubMed: 31829582DOI: 10.1021/jacs.9b10687 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.41 Å) |
Structure validation
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