5ZU2

Effect of mutation (R554A) on FAD modification in Aspergillus oryzae RIB40formate oxidase

5ZU2 の概要

• エントリーDOI: 10.2210/pdb5zu2/pdb
• 分子名称: formate oxidase, FLAVIN-ADENINE DINUCLEOTIDE, (4S)-2-METHYL-2,4-PENTANEDIOL, ... (5 entities in total)
• 機能のキーワード: formate oxidase, 8-formyl-fad, effect of mutation, oxidoreductase
• 由来する生物種: Aspergillus oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold)
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 195698.48

構造登録者

Mikami, B.,Uchida, H.,Doubayashi, D. (登録日: 2018-05-06, 公開日: 2019-05-22, 最終更新日: 2024-03-27)

主引用文献

Doubayashi, D.,Oki, M.,Mikami, B.,Uchida, H.
The microenvironment surrounding FAD mediates its conversion to 8-formyl-FAD in Aspergillus oryzae RIB40 formate oxidase.
J.Biochem., 166:67-75, 2019

PubMed Abstract: Aspergillus oryzae RIB40 formate oxidase has Arg87 and Arg554 near the formyl group and O(4) atom of 8-formyl-flavin adenine dinucleotide (FAD), respectively, with Asp396 neighbouring Arg554. Herein, we probed the roles of these three residues in modification of FAD to 8-formyl-FAD. Replacement of Arg87 or Arg554 with Lys or Ala decreased and abolished the modification, respectively. Replacement of Asp396 with Ala or Asn lowered the modification rate. The observation of unusual effects of maintaining pH 7.0 on the modification in R87K, R554K and D396 variants indicates initial and subsequent processes with different pH dependencies. Comparison of the initial process at pH 4.5 and 7.0 suggests that the microenvironment around Arg87 and the protonation state of Asp396 affect the initial process in the native enzyme. Comparison of the crystal structures of native and R554 variants showed that the replacements had minimal effect on catalytic site structure. The positively charged Arg87 might contribute to the formation of an anionic quinone-methide tautomer intermediate, while the positively charged Arg554, in collaboration with the negatively charged Asp396, might stabilize this intermediate and form a hydrogen bonding network with the N(5)/O(4) region, thereby facilitating efficient FAD modification.

PubMed: 30715389
DOI: 10.1093/jb/mvz009

実験手法

X-RAY DIFFRACTION (2.441 Å)