6A1B
Mandelate oxidase mutant-Y128F with 3,3,3-trifluoro-2,2-dihydroxypropanoic acid
Summary for 6A1B
| Entry DOI | 10.2210/pdb6a1b/pdb |
| Descriptor | 4-hydroxymandelate oxidase, FLAVIN MONONUCLEOTIDE, 3,3,3-trifluoro-2,2-dihydroxypropanoic acid, ... (4 entities in total) |
| Functional Keywords | fmn-dependent oxidase, flavoprotein |
| Biological source | Amycolatopsis orientalis (Nocardia orientalis) |
| Total number of polymer chains | 1 |
| Total formula weight | 40645.95 |
| Authors | |
| Primary citation | Lin, K.H.,Lyu, S.Y.,Yeh, H.W.,Li, Y.S.,Hsu, N.S.,Huang, C.M.,Wang, Y.L.,Shih, H.W.,Wang, Z.C.,Wu, C.J.,Li, T.L. Structural and chemical trapping of flavin-oxide intermediates reveals substrate-directed reaction multiplicity. Protein Sci., 29:1655-1666, 2020 Cited by PubMed Abstract: Though reactive flavin-N5/C4α-oxide intermediates can be spectroscopically profiled for some flavin-assisted enzymatic reactions, their exact chemical configurations are hardly visualized. Structural systems biology and stable isotopic labelling techniques were exploited to correct this stereotypical view. Three transition-like complexes, the α-ketoacid…N5-FMN complex (I), the FMN -N5-aloxyl-C'α -C4α zwitterion (II), and the FMN-N5-ethenol-N5-C4α-epoxide (III), were determined from mandelate oxidase (Hmo) or its mutant Y128F (monooxygenase) crystals soaked with monofluoropyruvate (a product mimic), establishing that N5 of FMN an alternative reaction center can polarize to an ylide-like mesomer in the active site. In contrast, four distinct flavin-C4α-oxide adducts (IV-VII) from Y128F crystals soaked with selected substrates materialize C4α of FMN an intrinsic reaction center, witnessing oxidation, Baeyer-Villiger/peroxide-assisted decarboxylation, and epoxidation reactions. In conjunction with stopped-flow kinetics, the multifaceted flavin-dependent reaction continuum is physically dissected at molecular level for the first time. PubMed: 32362037DOI: 10.1002/pro.3879 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.47 Å) |
Structure validation
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