6A01

The crystal structure of Mandelate oxidase Y128F with 3,3-difluoro-2,2-dihydroxy-3-phenylpropionic acid

6A01 の概要

• エントリーDOI: 10.2210/pdb6a01/pdb
• 分子名称: 4-hydroxymandelate oxidase, FLAVIN MONONUCLEOTIDE, 3,3-difluoro-2,2-dihydroxy-3-phenylpropanoic acid, ... (4 entities in total)
• 機能のキーワード: fmn-dependent oxidase, flavoprotein
• 由来する生物種: Amycolatopsis orientalis (Nocardia orientalis)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 40704.06

構造登録者

Li, T.L.,Lin, K.H. (登録日: 2018-06-05, 公開日: 2019-06-19, 最終更新日: 2023-11-22)

主引用文献

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

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: 32362037
DOI: 10.1002/pro.3879

実験手法

X-RAY DIFFRACTION (1.868 Å)