5YS1

Crystal structure of Multicopper Oxidase CueO G304K mutant

5YS1 の概要

• エントリーDOI: 10.2210/pdb5ys1/pdb
• 分子名称: Blue copper oxidase CueO, COPPER (II) ION (3 entities in total)
• 機能のキーワード: multicopper oxidase, cu-binding protein, cueo mutant, metal binding protein
• 由来する生物種: Escherichia coli K12
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 56885.75

構造登録者

Wang, H.Q.,Liu, X.Q.,Zhao, J.T.,Yue, Q.X.,Yan, Y.H.,Dong, Y.H.,Fan, Y.L.,Tian, J.,Wu, N.F.,Gong, Y. (登録日: 2017-11-12, 公開日: 2018-10-17, 最終更新日: 2023-11-22)

主引用文献

Wang, H.,Liu, X.,Zhao, J.,Yue, Q.,Yan, Y.,Gao, Z.,Dong, Y.,Zhang, Z.,Fan, Y.,Tian, J.,Wu, N.,Gong, Y.
Crystal structures of multicopper oxidase CueO G304K mutant: structural basis of the increased laccase activity
Sci Rep, 8:14252-14252, 2018

PubMed Abstract: The multicopper oxidase CueO is involved in copper homeostasis and copper (Cu) tolerance in Escherichia coli. The laccase activity of CueO G304K mutant is higher than wild-type CueO. To explain this increase in activity, we solved the crystal structure of G304K mutant at 1.49 Å. Compared with wild-type CueO, the G304K mutant showed dramatic conformational changes in methionine-rich helix and the relative regulatory loop (R-loop). We further solved the structure of Cu-soaked enzyme, and found that the addition of Cu ions induced further conformational changes in the R-loop and methionine-rich helix as a result of the new Cu-binding sites on the enzyme's surface. We propose a mechanism for the enhanced laccase activity of the G304K mutant, where movements of the R-loop combined with the changes of the methionine-rich region uncover the T1 Cu site allowing greater access of the substrate. Two of the G304K double mutants showed the enhanced or decreased laccase activity, providing further evidence for the interaction between the R-loop and the methionine-rich region. The cuprous oxidase activity of these mutants was about 20% that of wild-type CueO. These structural features of the G304K mutant provide clues for designing specific substrate-binding mutants in the biotechnological applications.

PubMed: 30250139
DOI: 10.1038/s41598-018-32446-7

実験手法

X-RAY DIFFRACTION (1.49 Å)