8U8T

Y229F/V290N Streptomyces coelicolor Laccase

8U8T の概要

• エントリーDOI: 10.2210/pdb8u8t/pdb
• 分子名称: Copper oxidase, COPPER (II) ION, HYDROXIDE ION, ... (9 entities in total)
• 機能のキーワード: laccase, copper, oxidoreductase
• 由来する生物種: Streptomyces coelicolor
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 117942.01

構造登録者

Wang, J.-X.,Lu, Y. (登録日: 2023-09-18, 公開日: 2023-11-15, 最終更新日: 2023-12-27)

主引用文献

Wang, J.X.,Vilbert, A.C.,Cui, C.,Mirts, E.N.,Williams, L.H.,Kim, W.,Jessie Zhang, Y.,Lu, Y.
Increasing Reduction Potentials of Type 1 Copper Center and Catalytic Efficiency of Small Laccase from Streptomyces coelicolor through Secondary Coordination Sphere Mutations.
Angew.Chem.Int.Ed.Engl., 62:e202314019-e202314019, 2023

PubMed Abstract: The key to type 1 copper (T1Cu) function lies in the fine tuning of the Cu reduction potential (E°' ) to match those of its redox partners, enabling efficient electron transfer in a wide range of biological systems. While the secondary coordination sphere (SCS) effects have been used to tune E°' in azurin over a wide range, these principles are yet to be generalized to other T1Cu-containing proteins to tune catalytic properties. To this end, we have examined the effects of Y229F, V290N and S292F mutations around the T1Cu of small laccase (SLAC) from Streptomyces coelicolor to match the high E°' of fungal laccases. Using ultraviolet-visible absorption and electron paramagnetic resonance spectroscopies, together with X-ray crystallography and redox titrations, we have probed the influence of SCS mutations on the T1Cu and corresponding E°' . While minimal and small E°' increases are observed in Y229F- and S292F-SLAC, the V290N mutant exhibits a major E°' increase. Moreover, the influence of these mutations on E°' is additive, culminating in a triple mutant Y229F/V290N/S292F-SLAC with the highest E°' of 556 mV vs. SHE reported to date. Further activity assays indicate that all mutants retain oxygen reduction reaction activity, and display improved catalytic efficiencies (k /K ) relative to WT-SLAC.

PubMed: 37926680
DOI: 10.1002/anie.202314019

実験手法

X-RAY DIFFRACTION (2.9 Å)