7PXM

X-ray structure of LPMO at 1.45x10^6 Gy

Summary for 7PXM

• Entry DOI: 10.2210/pdb7pxm/pdb
• Related: 7PXI 7PXJ 7PXK 7PXL
• Descriptor: Auxiliary activity 9, COPPER (II) ION, 2-acetamido-2-deoxy-beta-D-glucopyranose, ... (4 entities in total)
• Functional Keywords: lytic polysaccharide monooxygenase, metalloenzyme, aa9, oxidoreductase
• Biological source: Lentinus similis (Panus similis)
• Total number of polymer chains: 1
• Total formula weight: 25557.60

Authors

Tandrup, T.,Lo Leggio, L. (deposition date: 2021-10-08, release date: 2022-08-24, Last modification date: 2024-01-31)

Primary citation

Tandrup, T.,Muderspach, S.J.,Banerjee, S.,Santoni, G.,Ipsen, J.O.,Hernandez-Rollan, C.,Norholm, M.H.H.,Johansen, K.S.,Meilleur, F.,Lo Leggio, L.
Changes in active-site geometry on X-ray photoreduction of a lytic polysaccharide monooxygenase active-site copper and saccharide binding.
Iucrj, 9:666-681, 2022

PubMed Abstract: The recently discovered lytic polysaccharide monooxygenases (LPMOs) are Cu-containing enzymes capable of degrading polysaccharide substrates oxidatively. The generally accepted first step in the LPMO reaction is the reduction of the active-site metal ion from Cu to Cu. Here we have used a systematic diffraction data collection method to monitor structural changes in two AA9 LPMOs, one from (AA9_A) and one from (AA9_A), as the active-site Cu is photoreduced in the X-ray beam. For AA9_A, the protein produced in two different recombinant systems was crystallized to probe the effect of post-translational modifications and different crystallization conditions on the active site and metal photoreduction. We can recommend that crystallographic studies of AA9 LPMOs wishing to address the Cu form use a total X-ray dose below 3 × 10 Gy, while the Cu form can be attained using 1 × 10 Gy. In all cases, we observe the transition from a hexa-coordinated Cu site with two solvent-facing ligands to a T-shaped geometry with no exogenous ligands, and a clear increase of the θ parameter and a decrease of the θ parameter by averages of 9.2° and 8.4°, respectively, but also a slight increase in θ. Thus, the θ and θ parameters are helpful diagnostics for the oxidation state of the metal in a His-brace protein. On binding of cello-oligosaccharides to AA9_A, regardless of the production source, the θ parameter increases, making the Cu site less planar, while the active-site Tyr-Cu distance decreases reproducibly for the Cu form. Thus, the θ increase found on copper reduction may bring AA9_A closer to an oligosaccharide-bound state and contribute to the observed higher affinity of reduced AA9_A for cellulosic substrates.

PubMed: 36071795
DOI: 10.1107/S2052252522007175

Experimental method

X-RAY DIFFRACTION (1.3 Å)