5n2i

F420:NADPH oxidoreductase from Thermobifida fusca with NADP+ bound

spinqualitylabelsall models 5n2i, resolution 1.80Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5n2i is a 4 chain structure with sequence from Thermobifida fusca YX. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.8Å Ligands: GOL, NAP Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function Q47RA9_THEFY Publication Abstract from PubMed F420H2-dependent enzymes reduce a wide range of substrates that are otherwise recalcitrant to enzyme-catalyzed reduction, and their potential for applications in biocatalysis has attracted increasingly attention. Thermobifida fusca is a moderately thermophilic bacterium and holds high biocatalytic potential as a source for several highly thermostable enzymes. We report here on the isolation and characterization of a thermostable F420:NADPH oxidoreductase (Tfu-FNO) from T. fusca, being the first F420-dependent enzyme described from this bacterium. Tfu-FNO was heterologously expressed in Escherichia coli, yielding up to 200 mg recombinant enzyme per liter of culture. We found that Tfu-FNO is highly thermostable, reaching its highest activity at 65 degrees C and that Tfu-FNO is likely to act in vivo as an F420 reductase at the expense of NADPH, similar to its counterpart in Streptomyces griseus We obtained the crystal structure of FNO in complex with NADP+ at 1.8 A resolution, providing the first bacterial FNO structure. The overall architecture and NADP+-binding site of Tfu-FNO were highly similar to those of the Archaeoglobus fulgidus FNO (Af-FNO). The active site is located in a hydrophobic pocket between an N-terminal dinucleotide-binding domain and a smaller C-terminal do-main. Residues interacting with the 2'-phosphate of NADP+ were probed by targeted mutagenesis, indicating that Thr28, Ser50, Arg51, and Arg55 are important for discriminating between NADP+ and NAD+. Interestingly, a T28A mutant increased the kinetic efficiency more than three-fold as compared with the wild-type enzyme when NADH is the substrate. The biochemical and structural data presented here provide crucial insights into the molecular recognition of the two cofactors, F420 and NAD(P)H by FNO. Isolation and characterization of a thermostable F420:NADPH oxidoreductase from Thermobifida fusca.,Kumar H, Nguyen QT, Binda C, Mattevi A, Fraaije MW J Biol Chem. 2017 Apr 14. pii: jbc.M117.787754. doi: 10.1074/jbc.M117.787754. PMID:28411200 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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