Identification of a Baeyer–Villiger monooxygenase sequence motif

Fraaije, Marco W.; Kamerbeek, Nanne M.; Berkel, Willem J.H. van; Janssen, Dick B.

doi:10.1016/s0014-5793(02)02623-6

Cited by 202 publications

(191 citation statements)

References 34 publications

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“…2). A DALI (16) search shows that there are no other three-dimensional protein structures that have an inserted subdomain of this topology, which seems, therefore, characteristic of the Baeyer-Villiger monooxygenase class of enzymes (27,28).…”

Section: Resultsmentioning

confidence: 99%

“…As indicated by the threedimensional structures of many flavoenzymes (24), hydride transfer is brought about by positioning the nicotinamide adjacent to the flavin so that the two rings overlap each other. In the case of PAMO, this process must necessarily involve a movement of the Arg-337 side chain, which must shift away from the (27), are outlined in red. His-173 is a strictly conserved residue of the fingerprint motif that has been shown to be crucial for catalysis.…”

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

“…The amino acid sequences of the Baeyer-Villiger monooxygenases are characterized by the conserved sequence motif FXGXXXHXXXW (27), which in PAMO corresponds to residues 167-177 (Figs. 2, 3, and 6).…”

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

“…Mutagenesis studies have shown that the strictly conserved His residue belonging to the fingerprint sequence (His-173 in PAMO) is crucial for catalysis and FAD binding. In cyclohexanone monooxygenase (33), the replacement of this His residue with Gln causes a 10-fold reduction in activity, whereas in 4-hydroxyacetophenone monooxygenase (27), the His3Ala mutation makes the enzyme inactive. In the crystal structure, His-173 is fully solvent exposed being part of the domain linker region at 16 Å from the flavin and at Ͼ10 Å from the NADP-binding site (Figs.…”

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

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Crystal structure of a Baeyer–Villiger monooxygenase

Malito

Alfieri²,

Fraaije³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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305

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Flavin-containing Baeyer-Villiger monooxygenases employ NADPH and molecular oxygen to catalyze the insertion of an oxygen atom into a carbon-carbon bond of a carbonylic substrate. These enzymes can potentially be exploited in a variety of biocatalytic applications given the wide use of Baeyer-Villiger reactions in synthetic organic chemistry. The catalytic activity of these enzymes involves the formation of two crucial intermediates: a flavin peroxide generated by the reaction of the reduced flavin with molecular oxygen and the ''Criegee'' intermediate resulting from the attack of the flavin peroxide onto the substrate that is being oxygenated. The crystal structure of phenylacetone monooxygenase, a Baeyer-Villiger monooxygenase from the thermophilic bacterium Thermobifida fusca, exhibits a two-domain architecture resembling that of the disulfide oxidoreductases. The active site is located in a cleft at the domain interface. An arginine residue lays above the flavin ring in a position suited to stabilize the negatively charged flavin-peroxide and Criegee intermediates. This amino acid residue is predicted to exist in two positions; the ''IN'' position found in the crystal structure and an ''OUT'' position that allows NADPH to approach the flavin to reduce the cofactor. Domain rotations are proposed to bring about the conformational changes involved in catalysis. The structural studies highlight the functional complexity of this class of flavoenzymes, which coordinate the binding of three substrates (molecular oxygen, NADPH, and phenylacetone) in proximity of the flavin cofactor with formation of two distinct catalytic intermediates.flavoenzyme ͉ mechanism of catalysis ͉ biocatalysis ͉ Baeyer-Villiger reaction ͉ crystallography A t the end of the 19th century, Baeyer and Villiger (1) discovered that cyclic ketones react with oxidants, such as peroxymonosulfuric acid, to yield lactones. The mechanism of the Baeyer-Villiger reaction involves a nucleophilic attack of a peroxide to the ketone reagent to generate the so-called ''Criegee'' intermediate ( Fig. 1), which is followed by an intramolecular rearrangement that leads to the migration of an alkyl group to an oxygen atom, generating the lactone product. Baeyer-Villiger reactions are of enormous value in synthetic organic chemistry, and the number of their applications is countless.Several microorganisms produce enzymes capable to catalyze Baeyer-Villiger reactions. These proteins are extensively studied for their exploitation in biocatalytic applications (2, 3). This interest follows the problems related to the toxicity and instability of the oxidizing reactants that are currently being used in chemical processes. In addition, enzymatic reactions exhibit a superior degree of enantio-and regioselectivity. Baeyer-Villiger monooxygenases are classified depending on the nature of their flavin cofactor (4). The type I enzymes are the most extensively investigated (5). They are FAD-dependent proteins that use NADPH and molecular oxygen to insert an oxygen atom into th...

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Section: Resultsmentioning

confidence: 99%

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

Section: -Hydroxyacetophenone Monooxygenasementioning

confidence: 99%

See 2 more Smart Citations

Crystal structure of a Baeyer–Villiger monooxygenase

Malito

Alfieri²,

Fraaije³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

275

305

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“…When these two protein structures are superimposed (Cα backbone) in spite of only 23.4% sequence identity, they present a highly conserved overall structure. The primary amino acid sequence of hFMO3 was aligned with the sequences of these two proteins and the FMO-identifying sequence motif together with the two Rossmann-fold motifs (Fraaije et al, 2002) were identified and are shown in Fig. 1.…”

Section: Sequence Alignment and Homology Modelingmentioning

confidence: 99%

Human flavin-containing monooxygenase 3: Structural mapping of gene polymorphisms and insights into molecular basis of drug binding

Gao

Catucci

Nardo

et al. 2016

Gene

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Human hepatic flavin-containing monooxygenase 3 (hFMO3) catalyses the monooxygenation of carbon-bound reactive heteroatoms and plays an important role in the metabolism of drugs and xenobiotics. Although numerous hFMO3 allelic variants have been identified in patients and their biochemical properties well-characterised in vitro, the molecular mechanisms underlying loss-offunction mutations have still not been elucidated due to lack of detailed structural information of hFMO3. Therefore, in this work a 3D structural model of hFMO3 was generated by homology modeling, evaluated by a variety of different bioinformatics tools, refined by molecular dynamics simulations and further assessed based on in vitro biochemical data. The molecular dynamics simulation results highlighted 4 flexible regions of the protein with some of them overlapping the data from trypsin digest. This was followed by structural mapping of 12 critical polymorphic variants and molecular docking experiments with five different known substrates/drugs of hFMO3 namely, benzydamine, sulindac sulphide, tozasertib, methimazole and trimethylamine. Localisation of these mutations on the hFMO3 model provided a structural explanation for their observed biological effects and docked models of hFMO3-drug complexes gave insights into their binding mechanis m demonstrating that nitrogen-and sulfur-containing substrates interact with the isoalloxazine ring through Pi-Cation interaction and Pi-Sulfur interactions, respectively. Finally, the data presented give insights into the drug binding mechanism of hFMO3 which could be valuable not only for screening of new chemical entities but more significantly for designing of novel inhibitors of this important Phase I drug metabolising enzyme.

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Flavoenzyme-Catalyzed Oxygenations and Oxidations of Phenolic Compounds

Moonen

Fraaije

Rietjens

et al. 2002

Advanced Synthesis & Catalysis

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Flavin‐dependent monooxygenases and oxidases play an important role in the mineralization of phenolic compounds. Because of their exquisite regioselectivity and stereoselectivity, these enzymes are of interest for the biocatalytic production of fine chemicals and food ingredients. In our group, we have characterized several flavoenzymes that act on phenolic compounds, including 4‐hydroxybenzoate 3‐hydroxylase, 3‐hydroxyphenylacetate 6‐hydroxylase, 4‐hydroxybenzoate 1‐hydroxylase (decarboxylating), hydroquinone hydroxylase, 2‐hydroxybiphenyl 3‐monooxygenase, phenol hydroxylase, 4‐hydroxyacetophenone monooxygenase and vanillyl‐alcohol oxidase. The catalytic properties of these enzymes are reviewed here, together with insights obtained from site‐directed and random mutagenesis.

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Identification of a Baeyer–Villiger monooxygenase sequence motif

Cited by 202 publications

References 34 publications

Crystal structure of a Baeyer–Villiger monooxygenase

Crystal structure of a Baeyer–Villiger monooxygenase

Human flavin-containing monooxygenase 3: Structural mapping of gene polymorphisms and insights into molecular basis of drug binding

Flavoenzyme-Catalyzed Oxygenations and Oxidations of Phenolic Compounds

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