Structural perturbations of substrate binding and oxidation state changes in a lytic polysaccharide monooxygenase

• Published in: Journal of biological inorganic chemistry Vol. 27; no. 8; pp. 705 - 713
• Main Authors: Walton, Paul H., Davies, Gideon J.
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.12.2022; Springer Nature B.V
• Subjects: Biochemistry; Biomedical and Life Sciences; Catalytic Domain; Enzymes; Inorganic chemistry; Intermediates; Life Sciences; Microbiology; Mixed Function Oxygenases - chemistry; Monooxygenase; Original Paper; Oxidation; Oxidation-Reduction; Polysaccharides; Polysaccharides - metabolism; Statistics; Substrate Specificity; Tryptophan; Tryptophan - metabolism; Errors; Copper; LPMO; Protein structure
• ISSN: 1432-1327; 0949-8257; 1432-1327
• DOI: 10.1007/s00775-022-01966-z

LPMOs are enzymes which catalyse the oxidation of a C-H bond within polysaccharides, leading to their oxidative cleavage. To achieve this, LPMOs employ highly reactive oxidising intermediates, the generation of which is likely coupled to substrate binding to the enzyme. The nature of this coupling is unknown. Here we report a statistical comparison for four three-dimensional structures of an AA9 LPMO crystallised in the same space group but in different oxidation and substrate-binding states, to determine which significant structural perturbations occur at the enzyme upon either oxidation state change or the binding of substrate. In a novel step, we determine the global random error associated with the positional coordinates of atoms using the method of moments to ascertain the statistical estimators of Gaussian distributions of pairwise RMS differences between individual atoms in different structures. The results show that a change in the oxidation state of the copper leads to no significant structural changes, and that binding of the substrate leads to a single change in the conformation of a tryptophan residue. This tryptophan has previously been identified as part of a charge transfer pathway between the active site and the external surface of the protein, and the structural change identified herein may be part of the substrate-enzyme coupling mechanism. Graphical abstract