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Title: The oxygenating constituent of 3,6-diketocamphane monooxygenase from the CAM plasmid of Pseudomonas putida: the first crystal structure of a type II Baeyer–Villiger monooxygenase

The first crystal structure of a type II Baeyer–Villiger monooxygenase reveals a different ring orientation of its FMN cofactor compared with other related bacterial luciferase-family enzymes. The three-dimensional structures of the native enzyme and the FMN complex of the overexpressed form of the oxygenating component of the type II Baeyer–Villiger 3,6-diketocamphane monooxygenase have been determined to 1.9 Å resolution. The structure of this dimeric FMN-dependent enzyme, which is encoded on the large CAM plasmid of Pseudomonas putida, has been solved by a combination of multiple anomalous dispersion from a bromine crystal soak and molecular replacement using a bacterial luciferase model. The orientation of the isoalloxazine ring of the FMN cofactor in the active site of this TIM-barrel fold enzyme differs significantly from that previously observed in enzymes of the bacterial luciferase-like superfamily. The Ala77 residue is in a cis conformation and forms a β-bulge at the C-terminus of β-strand 3, which is a feature observed in many proteins of this superfamily.
Authors:
; ; ; ; ; ; ; ; ;  [1] ;  [2] ; ;  [3] ; ; ;  [4] ;  [5] ;  [1]
  1. University of Exeter, Stocker Road, Exeter EX4 4QD (United Kingdom)
  2. National Research Council Canada, 6100 Royalmount Avenue, Montreal, QC H4P 2R2 (Canada)
  3. Kansai University (Japan)
  4. Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
  5. European Molecular Biology Laboratory (EMBL), Hamburg Outstation, Notkestrasse 85, 22607 Hamburg (Germany)
Publication Date:
OSTI Identifier:
22515196
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 11; Other Information: PMCID: PMC4631483; PMID: 26527149; PUBLISHER-ID: tz5084; OAI: oai:pubmedcentral.nih.gov:4631483; Copyright (c) Isupov et al. 2015; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; PROTEIN STRUCTURE; RINGS; THREE-DIMENSIONAL CALCULATIONS