6VBJ
CRYSTAL STRUCTURE OF THE HYBRID C-TERMINAL DOMAIN OF ENZYME I OF THE BACTERIAL PHOSPHOTRANSFERASE SYSTEM FORMED BY HYBRIDIZING THE SCAFFOLD OF THE THERMOANAEROBACTER TENGCONGENSIS ENZYME WITH THE ACTIVE SITE LOOPS FROM THE ESCHERICHIA COLI ENZYME
Summary for 6VBJ
| Entry DOI | 10.2210/pdb6vbj/pdb |
| Descriptor | Phosphoenolpyruvate-protein phosphotransferase (2 entities in total) |
| Functional Keywords | phosphoenolpyruvate-protein phosphotransferase ptsi, hybrid, bacterial phosphotransferase, transferase |
| Biological source | Caldanaerobacter subterraneus subsp. tengcongensis |
| Total number of polymer chains | 2 |
| Total formula weight | 69782.79 |
| Authors | Stewart Jr., C.E. (deposition date: 2019-12-19, release date: 2020-06-17, Last modification date: 2023-10-11) |
| Primary citation | Dotas, R.R.,Nguyen, T.T.,Stewart Jr., C.E.,Ghirlando, R.,Potoyan, D.A.,Venditti, V. Hybrid Thermophilic/Mesophilic Enzymes Reveal a Role for Conformational Disorder in Regulation of Bacterial Enzyme I. J.Mol.Biol., 432:4481-4498, 2020 Cited by PubMed Abstract: Conformational disorder is emerging as an important feature of biopolymers, regulating a vast array of cellular functions, including signaling, phase separation, and enzyme catalysis. Here we combine NMR, crystallography, computer simulations, protein engineering, and functional assays to investigate the role played by conformational heterogeneity in determining the activity of the C-terminal domain of bacterial Enzyme I (EIC). In particular, we design chimeric proteins by hybridizing EIC from thermophilic and mesophilic organisms, and we characterize the resulting constructs for structure, dynamics, and biological function. We show that EIC exists as a mixture of active and inactive conformations and that functional regulation is achieved by tuning the thermodynamic balance between active and inactive states. Interestingly, we also present a hybrid thermophilic/mesophilic enzyme that is thermostable and more active than the wild-type thermophilic enzyme, suggesting that hybridizing thermophilic and mesophilic proteins is a valid strategy to engineer thermostable enzymes with significant low-temperature activity. PubMed: 32504625DOI: 10.1016/j.jmb.2020.05.024 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
Download full validation report
