9NJI

M298L Streptomyces coelicolor Laccase

Summary for 9NJI

• Entry DOI: 10.2210/pdb9nji/pdb
• Descriptor: Copper oxidase, COPPER (II) ION, HYDROXIDE ION, ... (6 entities in total)
• Functional Keywords: laccase, copper, oxidoreductase
• Biological source: Streptomyces coelicolor
• Total number of polymer chains: 1
• Total formula weight: 38945.02

Authors

Wang, J.-X.,Lu, Y. (deposition date: 2025-02-27, release date: 2025-06-04, Last modification date: 2025-07-02)

Primary citation

Wang, J.X.,Vilbert, A.C.,Williams, L.H.,Mirts, E.N.,Cui, C.,Lu, Y.
Unexpected effect of an axial ligand mutation in the type 1 copper center in small laccase: structure-based analyses and engineering to increase reduction potential and activity.
Chem Sci, 16:11339-11346, 2025

PubMed Abstract: Type 1 copper (T1Cu) centers are crucial in biological electron transfer (ET) processes, exhibiting a wide range of reduction potentials to match their redox partners and optimize ET rates. While tuning in mononuclear T1Cu proteins like azurin has been successful, it is more difficult for multicopper oxidases. Specifically, while replacing axial methionine to leucine in azurin increased its by ∼100 mV, the corresponding M298L mutation in small laccase from (SLAC) unexpectedly decreased its by 12 mV. X-ray crystallography revealed two axial water molecules in M298L-SLAC, leading to the decrease of due to decreased hydrophobicity. Structural alignment and molecular dynamics simulation indicated a key difference in T1Cu axial loop position, leading to the different outcome upon methionine to leucine mutation. Based on structural analyses, we introduced additional F195L and I200F mutations, leading to partial removal of axial waters, a 122-mV increase in , and a 7-fold increase in / from M298L-SLAC. These findings highlight the complexity of tuning in multicopper oxidases and provide valuable insights into how structure-based protein engineering can contribute to the broader understanding of T1Cu center, and reactivity tuning for applications, such as in solar energy transfer, fuel cells, and bioremediation.

PubMed: 40438172
DOI: 10.1039/d5sc02177d

Experimental method

X-RAY DIFFRACTION (2.6 Å)