3PL8
Pyranose 2-oxidase H167A complex with 3-deoxy-3-fluoro-beta-D-glucose
Summary for 3PL8
| Entry DOI | 10.2210/pdb3pl8/pdb |
| Related | 1TT0 2IGO |
| Descriptor | Pyranose 2-oxidase, FLAVIN-ADENINE DINUCLEOTIDE, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (5 entities in total) |
| Functional Keywords | substrate complex, h167a mutant, homotetramer, gmc oxidoreductase, phbh fold, rossmann domain, oxidoreductase, oxidase, fad cofactor, pyranose oxidation |
| Biological source | Trametes ochracea (White-rot fungus) |
| Total number of polymer chains | 1 |
| Total formula weight | 70509.64 |
| Authors | |
| Primary citation | Tan, T.C.,Haltrich, D.,Divne, C. Regioselective control of Beta-d-glucose oxidation by pyranose 2-oxidase is intimately coupled to conformational degeneracy J.Mol.Biol., 409:588-600, 2011 Cited by PubMed Abstract: Trametes multicolor pyranose 2-oxidase (P2O) is a flavoprotein oxidase that oxidizes d-glucose at C2 to 2-keto-d-glucose by a highly regioselective mechanism. In this work, fluorinated sugar substrates were used as mechanistic probes to investigate the basis of regioselectivity in P2O. Although frequently used to study the mechanisms of glycoside hydrolases, our work provides the first example of applying these probes to sugar oxidoreductases. Our previous structure of the P2O mutant H167A in complex with the slow substrate 2-deoxy-2-fluoro-d-glucose showed a substrate-binding mode compatible with oxidation at C3. To accommodate the sugar, a gating segment, (454)FSY(456), in the substrate recognition loop partly unfolded to create a spacious and more polar active site that is distinct from the closed state of P2O. The crystal structure presented here shows that the preferred C2 oxidation where an ordered complex of P2O H167A with 3-deoxy-3-fluoro-d-glucose at 1.35 Å resolution was successfully trapped. In this semi-open C2-oxidation complex, the substrate recognition loop tightens to form an optimized substrate complex stabilized by interactions between Asp452 and glucose O4, as well as Tyr456 and the glucose O6 group, interactions that are not possible when glucose is positioned for oxidation at C3. The different conformations of the (454)FSY(456) gating segment in the semi-open and closed states induce backbone and side-chain movements of Thr169 and Asp452 that add further differential stabilization to the individual states. We expect the semi-open state (C2-oxidation state) and closed state to be good approximations of the active-site structure during the reductive half-reaction (sugar oxidation) and oxidative half-reaction (O(2) reduction). PubMed: 21515286DOI: 10.1016/j.jmb.2011.04.019 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.35 Å) |
Structure validation
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