7PYL

Structure of an LPMO (expressed in E.coli) at 1.49x10^4 Gy

Summary for 7PYL

• Entry DOI: 10.2210/pdb7pyl/pdb
• Related: 7PQR 7PXI 7PXJ 7PXK 7PXL 7PXM 7PXN 7PXR 7PXS 7PXT 7PXU 7PXV 7PXW 7PYD 7PYE 7PYF 7PYG 7PYH 7PYI
• Descriptor: Auxiliary activity 9, COPPER (II) ION, ACETATE ION, ... (5 entities in total)
• Functional Keywords: lytic polysaccharide monooxygenase, metalloenzyme, copper, auxiliary activity, oxidoreductase
• Biological source: Lentinus similis (Panus similis)
• Total number of polymer chains: 1
• Total formula weight: 25766.67

Authors

Tandrup, T.,Muderspach, S.J.,Banerjee, S.,Ipsen, J.O.,Rollan, C.H.,Norholm, M.H.H.,Johansen, K.S.,Lo Leggio, L. (deposition date: 2021-10-10, release date: 2022-08-24, Last modification date: 2024-11-13)

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 (1.7 Å)