9TCZ

Structure of an LPMO expressed in E.coli (LsAA9A) at 6.40x10^4 Gy

This is a non-PDB format compatible entry.

Summary for 9TCZ

• Entry DOI: 10.2210/pdb9tcz/pdb
• Related PRD ID: PRD_900021
• Descriptor: Auxiliary activity 9, beta-D-glucopyranose-(1-4)-beta-D-glucopyranose-(1-4)-beta-D-glucopyranose, ACETATE ION, ... (7 entities in total)
• Functional Keywords: lytic polysaccharide monooxygenase, copper-binding enzyme, aa9 family, oxidoreductase
• Biological source: Panus similis
• Total number of polymer chains: 1
• Total formula weight: 26688.22

Authors

Wei, Q.,Huang, Z.,Lo Leggio, L. (deposition date: 2025-11-22, release date: 2026-08-05, Last modification date: 2026-08-12)

Primary citation

Huang, Z.,Wei, Q.,Nan, J.,Norholm, M.H.H.,Liu, Z.,Hernandez-Rollan, C.,Johansen, K.S.,Lo Leggio, L.
Experimental estimation of copper-site geometry reproducibility in biologically relevant redox and saccharide-bound states of a model lytic polysaccharide monooxygenase.
Acta Crystallogr D Struct Biol, 82:886-899, 2026

PubMed Abstract: Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes classified into the auxiliary activity (AA) families of the CAZy database. They oxidatively cleave glycosidic bonds in recalcitrant polysaccharides, playing a key role in biomass degradation and contributing to the virulence of some pathogens. The redox state of the active-site copper and its coordination geometry are central to LPMO catalysis, yet the subtle structural consequences of redox and substrate-binding transitions remain insufficiently resolved. In previous work, a comparative X-ray crystallographic analysis of a model AA9 LPMO (LsAA9A) was conducted under four distinct conditions at 100 K: Cu(II), Cu(I) and the corresponding saccharide-bound states, with the Cu(I) state generated by X-ray photoreduction. In this study, LsAA9A crystals were chemically reduced with or without saccharide substrate prior to low-dose X-ray data collection to minimize radiation damage. Copper-coordination distances and angles were determined precisely through triplicate structure determinations (each from an independent crystal) for each condition, revealing small but reproducible geometry differences across key states in the LPMO catalytic pathway. In order to identify the most significant differences, statistical evaluation using one-way analysis of variance (ANOVA), followed by Tukey-Kramer post hoc tests and pairwise t-tests, was carried out. Within the assumptions made, statistically significant differences in the coordinated Cu-His1 N and Cu-Tyr O distances, and in the coordination angles θ, θ and θ, are observed across the four states and are discussed in terms of the mechanism and in relation to our previous study. To complement cryogenic data, multi-crystal data sets at increasing X-ray dose were collected at room temperature to track photoreduction of the copper site, with the specific aim of detecting the transition of Cu(II) to fully reduced Cu(I). This could not finally be achieved due to the onset of global radiation damage; however, a subset of the reduction-linked geometric changes were detectable, indicating that a partial transition may have occurred.

PubMed: 42466526
DOI: 10.1107/S2059798326005966

Experimental method

X-RAY DIFFRACTION (2.06 Å)