7PXR

Room temperature structure of an LPMO.

7PXR の概要

• エントリーDOI: 10.2210/pdb7pxr/pdb
• 関連するPDBエントリー: 7PQR 7PXI 7PXJ 7PXK 7PXL 7PXM 7PXN
• 分子名称: Auxiliary activity 9, COPPER (II) ION, 2-acetamido-2-deoxy-beta-D-glucopyranose, ... (5 entities in total)
• 機能のキーワード: copper binding protein, beta-sandwich fold, auxillary, oxidoreductase
• 由来する生物種: Lentinus similis (Panus similis)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 26018.49

構造登録者

Tandrup, T.,Meilleur, F.,Ipsen, J.,Johansen, K.S.,Lo Leggio, L. (登録日: 2021-10-08, 公開日: 2022-08-24, 最終更新日: 2024-01-31)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.8 Å)