34 results on '"Mattevi, Andrea"'
Search Results
2. Same Substrate, Many Reactions: Oxygen Activation in Flavoenzymes.
- Author
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Romero E, Gómez Castellanos JR, Gadda G, Fraaije MW, and Mattevi A
- Subjects
- Hydrogen Peroxide metabolism, Mixed Function Oxygenases metabolism, Substrate Specificity, Enzymes metabolism, Flavins metabolism, Oxygen metabolism
- Abstract
Over time, organisms have evolved strategies to cope with the abundance of dioxygen on Earth. Oxygen-utilizing enzymes tightly control the reactions involving O
2 mostly by modulating the reactivity of their cofactors. Flavins are extremely versatile cofactors that are capable of undergoing redox reactions by accepting either one electron or two electrons, alternating between the oxidized and the reduced states. The physical and chemical principles of flavin-based chemistry have been investigated widely. In the following pages we summarize the state of the art on a key area of research in flavin enzymology: the molecular basis for the activation of O2 by flavin-dependent oxidases and monooxygenases. In general terms, oxidases use O2 as an electron acceptor to produce H2 O2 , while monooxygenases activate O2 by forming a flavin intermediate and insert an oxygen atom into the substrate. First, we analyze how O2 reaches the flavin cofactor embedded in the protein matrix through dedicated access pathways. Then we approach O2 activation from the perspective of the monooxygenases, their preferred intermediate, the C(4a)-(hydro)peroxyflavin, and the cases in which other intermediates have been described. Finally, we focus on understanding how the architectures developed in the active sites of oxidases promote O2 activation and which other factors operate in its reactivity.- Published
- 2018
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3. Structure and functionality in flavivirus NS-proteins: perspectives for drug design.
- Author
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Bollati M, Alvarez K, Assenberg R, Baronti C, Canard B, Cook S, Coutard B, Decroly E, de Lamballerie X, Gould EA, Grard G, Grimes JM, Hilgenfeld R, Jansson AM, Malet H, Mancini EJ, Mastrangelo E, Mattevi A, Milani M, Moureau G, Neyts J, Owens RJ, Ren J, Selisko B, Speroni S, Steuber H, Stuart DI, Unge T, and Bolognesi M
- Subjects
- Animals, Antiviral Agents chemistry, Antiviral Agents pharmacology, Biomedical Research organization & administration, Biomedical Research trends, Communicable Diseases, Emerging epidemiology, Communicable Diseases, Emerging virology, Drug Design, Enzyme Inhibitors chemistry, Enzyme Inhibitors pharmacology, Enzymes genetics, European Union, Flavivirus drug effects, Flavivirus Infections epidemiology, Flavivirus Infections virology, Humans, Viral Nonstructural Proteins antagonists & inhibitors, Viral Nonstructural Proteins genetics, Virus Replication drug effects, Enzymes chemistry, Enzymes metabolism, Flavivirus enzymology, Viral Nonstructural Proteins chemistry, Viral Nonstructural Proteins metabolism
- Abstract
Flaviviridae are small enveloped viruses hosting a positive-sense single-stranded RNA genome. Besides yellow fever virus, a landmark case in the history of virology, members of the Flavivirus genus, such as West Nile virus and dengue virus, are increasingly gaining attention due to their re-emergence and incidence in different areas of the world. Additional environmental and demographic considerations suggest that novel or known flaviviruses will continue to emerge in the future. Nevertheless, up to few years ago flaviviruses were considered low interest candidates for drug design. At the start of the European Union VIZIER Project, in 2004, just two crystal structures of protein domains from the flaviviral replication machinery were known. Such pioneering studies, however, indicated the flaviviral replication complex as a promising target for the development of antiviral compounds. Here we review structural and functional aspects emerging from the characterization of two main components (NS3 and NS5 proteins) of the flavivirus replication complex. Most of the reviewed results were achieved within the European Union VIZIER Project, and cover topics that span from viral genomics to structural biology and inhibition mechanisms. The ultimate aim of the reported approaches is to shed light on the design and development of antiviral drug leads., (Copyright 2009 Elsevier B.V. All rights reserved.)
- Published
- 2010
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4. Enzymes without borders: mobilizing substrates, delivering products.
- Author
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Forneris F and Mattevi A
- Subjects
- Alkyl and Aryl Transferases chemistry, Alkyl and Aryl Transferases metabolism, Amidohydrolases chemistry, Amidohydrolases metabolism, Binding Sites, Catalytic Domain, Cytosol enzymology, Cytosol metabolism, Diffusion, Hydrophobic and Hydrophilic Interactions, Hydroxysteroid Dehydrogenases chemistry, Hydroxysteroid Dehydrogenases metabolism, Intracellular Membranes enzymology, Membrane Proteins chemistry, Metalloproteases chemistry, Metalloproteases metabolism, Models, Chemical, Organelles enzymology, Peptidoglycan Glycosyltransferase chemistry, Peptidoglycan Glycosyltransferase metabolism, Enzymes chemistry, Enzymes metabolism, Intracellular Membranes metabolism, Lipid Bilayers metabolism, Membrane Proteins metabolism, Organelles metabolism
- Abstract
Many cellular reactions involve both hydrophobic and hydrophilic molecules that reside within the chemically distinct environments defined by the phospholipid-based membranes and the aqueous lumens of cytoplasm and organelles. Enzymes performing this type of reaction are required to access a lipophilic substrate located in the membranes and to catalyze its reaction with a polar, water-soluble compound. Here, we explore the different binding strategies and chemical tricks that enzymes have developed to overcome this problem. These reactions can be catalyzed by integral membrane proteins that channel a hydrophilic molecule into their active site, as well as by water-soluble enzymes that are able to capture a lipophilic substrate from the phospholipid bilayer. Many chemical and biological aspects of this type of enzymology remain to be investigated and will require the integration of protein chemistry with membrane biology.
- Published
- 2008
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5. New frontiers in structural flavoenzymology.
- Author
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De Colibus L and Mattevi A
- Subjects
- Catalysis, Flavin Mononucleotide biosynthesis, Flavin Mononucleotide chemistry, Flavin-Adenine Dinucleotide biosynthesis, Flavin-Adenine Dinucleotide chemistry, Histone Demethylases, Humans, In Vitro Techniques, Models, Molecular, Molecular Structure, Oxidoreductases, N-Demethylating chemistry, Oxidoreductases, N-Demethylating metabolism, Oxygen metabolism, Protein Conformation, Enzymes chemistry, Enzymes metabolism, Flavoproteins chemistry, Flavoproteins metabolism
- Abstract
During the past few years, there have been exciting developments in the field of flavoenzymology. New flavoenzymes have been discovered that are implicated in a variety of biological processes, including cell signaling, chromatin remodeling and cell development. The structures of several of these new flavoenzymes have been described, as exemplified by crystallographic analyses of MICAL, histone demethylase LSD1 and tryptophan dehalogenase. In addition, new structural information has revealed the evolutionary and mechanistic complexity of the enzymes of the riboflavin biosynthetic pathway. The integration of the enzymology data with crystallographic studies at atomic resolution is resulting in unprecedented insight into the chemical and geometric properties underlying flavoenzyme function.
- Published
- 2006
- Full Text
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6. To be or not to be an oxidase: challenging the oxygen reactivity of flavoenzymes.
- Author
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Mattevi A
- Subjects
- Enzymes chemistry, Flavoproteins chemistry, Models, Biological, Oxidoreductases chemistry, Oxygen chemistry, Enzymes metabolism, Flavins metabolism, Flavoproteins metabolism, Oxidoreductases metabolism, Oxygen metabolism
- Abstract
Flavin-dependent enzymes catalyse a wide range of reactions and, thereby, facilitate a variety of cellular processes. Among the properties that equip flavoenzymes with this chemical versatility is their reactivity towards oxygen, which shows huge variation among flavoproteins. A survey of known 3D structures of flavin-dependent oxidases and dehydrogenases and the correlation with their functional properties indicates that there are no structural rules that enable prediction of whether or how a flavoenzyme reacts with oxygen. Combinations of subtle factors such as dipole pre-organization, charge distribution, dynamics and solvation in the active centre determine the balance of interactions that control oxygen reactivity. The chemical basis of oxygen reactivity remains a puzzling problem and represents one of the challenging questions in modern flavoenzymology.
- Published
- 2006
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7. Three-Dimensional Structure of Human Monoamine Oxidase A (MAO A): Relation to the Structures of Rat MAO A and Human MAO B
- Author
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De Colibus, Luigi, Li, Min, Binda, Claudia, Lustig, Ariel, Edmondson, Dale E., Mattevi, Andrea, and Klinman, Judith P.
- Published
- 2005
8. Crystal Structure of a Baeyer-Villiger Monooxygenase
- Author
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Malito, Enrico, Alfieri, Andrea, Fraaije, Marco W., Mattevi, Andrea, and Matthews, Rowena G.
- Published
- 2004
9. Insights into the Mode of Inhibition of Human Mitochondrial Monoamine Oxidase B from High-Resolution Crystal Structures
- Author
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Binda, Claudia, Li, Min, Hubálek, Frantisek, Restelli, Nadia, Edmondson, Dale E., and Mattevi, Andrea
- Published
- 2003
10. Inversion of Stereospecificity of Vanillyl-Alcohol Oxidase
- Author
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Fraaije, Marco W., Ferrer, Miriam, and Mattevi, Andrea
- Published
- 2000
11. Crystal Structure of D-Amino Acid Oxidase: A Case of Active Site Mirror-Image Convergent Evolution with Flavocytochrome b2
- Author
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Mattevi, Andrea, Vanoni, Maria Antonietta, Todone, Flavia, Rizzi, Menico, Teplyakov, Alex, Coda, Alessandro, Bolognesi, Martino, and Curti, Bruno
- Published
- 1996
12. Precursor of ether phospholipids is synthesized by a flavoenzyme through covalent catalysis
- Author
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Nenci, Simone, Piano, Valentina, Rosati, Sara, Aliverti, Alessandro, Pandini, Vittorio, Fraaije, Marco W., Heck, Albert J. R., Edmondson, Dale E., and Mattevi, Andrea
- Published
- 2012
13. Evolution of enzyme functionality in the flavin-containing monooxygenases.
- Author
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Bailleul, Gautier, Yang, Guang, Nicoll, Callum R., Mattevi, Andrea, Fraaije, Marco W., and Mascotti, Maria Laura
- Subjects
MONOOXYGENASES ,ENZYMES ,XENOBIOTICS ,AMINO acids ,TETRAPODS - Abstract
Among the molecular mechanisms of adaptation in biology, enzyme functional diversification is indispensable. By allowing organisms to expand their catalytic repertoires and adopt fundamentally different chemistries, animals can harness or eliminate new-found substances and xenobiotics that they are exposed to in new environments. Here, we explore the flavin-containing monooxygenases (FMOs) that are essential for xenobiotic detoxification. Employing a paleobiochemistry approach in combination with enzymology techniques we disclose the set of historical substitutions responsible for the family's functional diversification in tetrapods. Remarkably, a few amino acid replacements differentiate an ancestral multi-tasking FMO into a more specialized monooxygenase by modulating the oxygenating flavin intermediate. Our findings substantiate an ongoing premise that enzymatic function hinges on a subset of residues that is not limited to the active site core. Detoxification enzymes are crucial for the survival of animals in new environments. Here, the authors study the molecular mechanism behind the catalytic diversification of a major family of tetrapod detoxification enzymes—the FMOs—during evolution. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
14. Multiple Pathways Guide Oxygen Diffusion into Flavoenzyme Active Sites
- Author
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Baron, Riccardo, Riley, Conor, Chenprakhon, Pirom, Thotsaporn, Kittisak, Winter, Remko T., Alfieri, Andrea, Forneris, Federico, van Berkels, Willem J. H., Chaiyen, Pimchai, Fraaije, Marco W., Mattevi, Andrea, McCammon, J. Andrew, and Eaton, William A.
- Published
- 2009
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15. Revealing the Moonlighting Role of NADP in the Structure of a Flavin-Containing Monooxygenase
- Author
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Alfieri, Andrea, Malito, Enrico, Orru, Roberto, Fraaije, Marco W., and Mattevi, Andrea
- Published
- 2008
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16. Structure of the Monooxygenase Component of a Two-Component Flavoprotein Monooxygenase
- Author
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Alfieri, Andrea, Fersini, Francesco, Ruangchan, Nantidaporn, Prongjit, Methinee, Chaiyen, Pimchai, and Mattevi, Andrea
- Published
- 2007
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17. Structure- and computational-aided engineering of an oxidase to produce isoeugenol from a lignin-derived compound.
- Author
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Guo, Yiming, Alvigini, Laura, Trajkovic, Milos, Alonso-Cotchico, Lur, Monza, Emanuele, Savino, Simone, Marić, Ivana, Mattevi, Andrea, and Fraaije, Marco W.
- Subjects
LIGNINS ,CATALYTIC activity ,ENZYMES ,ENGINEERING ,LIGNIN structure ,CHEMOSELECTIVITY ,EUGENOL - Abstract
Various 4-alkylphenols can be easily obtained through reductive catalytic fractionation of lignocellulosic biomass. Selective dehydrogenation of 4-n-propylguaiacol results in the formation of isoeugenol, a valuable flavor and fragrance molecule and versatile precursor compound. Here we present the engineering of a bacterial eugenol oxidase to catalyze this reaction. Five mutations, identified from computational predictions, are first introduced to render the enzyme more thermostable. Other mutations are then added and analyzed to enhance chemoselectivity and activity. Structural insight demonstrates that the slow catalytic activity of an otherwise promising enzyme variant is due the formation of a slowly-decaying covalent substrate-flavin cofactor adduct that can be remedied by targeted residue changes. The final engineered variant comprises eight mutations, is thermostable, displays good activity and acts as a highly chemoselective 4-n-propylguaiacol oxidase. We lastly use our engineered biocatalyst in an illustrative preparative reaction at gram-scale. Our findings show that a natural enzyme can be redesigned into a tailored biocatalyst capable of valorizing lignin-based monophenols. Lignin can be depolymerized into 4-alkylphenols by chemical means. Here the authors show a three-step computational-assisted enzyme engineering process to generate a biocatalyst for the conversion of lignin-derived 4-n-propylguaiacol into isoeugenol, a valuable compound. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
18. Atomic Structure of the Cubic Core of the Pyruvate Dehydrogenase Multienzyme Complex
- Author
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Mattevi, Andrea, Obmolova, Galya, Schulze, Egbert, Kalk, Kor H., Westphal, Adrie H., and De Kok, Arie
- Published
- 1992
19. Characterization of a thermostable flavin-containing monooxygenase from Nitrincola lacisaponensis (NiFMO).
- Author
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Lončar, Nikola, Fiorentini, Filippo, Bailleul, Gautier, Savino, Simone, Romero, Elvira, Mattevi, Andrea, and Fraaije, Marco W.
- Subjects
MONOOXYGENASES ,DRUG metabolism ,ENZYMES ,ORGANIC solvents ,ESCHERICHIA coli - Abstract
The flavin-containing monooxygenases (FMOs) play an important role in drug metabolism but they also have a high potential in industrial biotransformations. Among the hitherto characterized FMOs, there was no thermostable representative, while such biocatalyst would be valuable for FMO-based applications. Through a targeted genome mining approach, we have identified a gene encoding for a putative FMO from Nitrincola lacisaponensis, an alkaliphilic extremophile bacterium. Herein, we report the biochemical and structural characterization of this newly discovered bacterial FMO (NiFMO). NiFMO can be expressed as active and soluble enzyme at high level in Escherichia coli (90-100 mg/L of culture). NiFMO is relatively thermostable (melting temperature (T
m ) of 51 °C), displays high organic solvent tolerance, and accepts a broad range of substrates. The crystal structure of NiFMO was solved at 1.8 Å resolution, which allows future structure-based enzyme engineering. Altogether, NiFMO represents an interesting newly discovered enzyme with the appropriate features to develop into an industrially applied biocatalyst. [ABSTRACT FROM AUTHOR]- Published
- 2019
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20. Kinetic Resolution of <italic>sec</italic>‐Thiols by Enantioselective Oxidation with Rationally Engineered 5‐(Hydroxymethyl)furfural Oxidase.
- Author
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Pickl, Mathias, Swoboda, Alexander, Faber, Kurt, Winkler, Christoph K., Romero, Elvira, Fraaije, Marco W., Binda, Claudia, and Mattevi, Andrea
- Subjects
THIOLS ,KINETIC resolution ,HYDROXYMETHYLFURFURAL ,ENANTIOSELECTIVE catalysis ,OXIDATION ,BIOCATALYSIS ,FLAVOPROTEINS ,REACTIVITY (Chemistry) - Abstract
Abstract: Various flavoprotein oxidases were recently shown to oxidize primary thiols. Herein, this reactivity is extended to
sec ‐thiols by using structure‐guided engineering of 5‐(hydroxymethyl)furfural oxidase (HMFO). The variants obtained were employed for the oxidative kinetic resolution of racemicsec ‐thiols, thus yielding the corresponding thioketones and nonreactedR ‐configured thiols with excellent enantioselectivities (E ≥200). The engineering strategy applied went beyond the classic approach of replacing bulky amino acid residues with smaller ones, as the active site was additionally enlarged by a newly introduced Thr residue. This residue established a hydrogen‐bonding interaction with the substrates, as verified in the crystal structure of the variant. These strategies unlocked HMFO variants for the enantioselective oxidation of a range ofsec ‐thiols. [ABSTRACT FROM AUTHOR]- Published
- 2018
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21. Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase.
- Author
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Romero, Elvira, Castellanos, J. Rubén Gómez, Mattevi, Andrea, and Fraaije, Marco W.
- Subjects
CRYSTAL structure ,CYCLOHEXANONES ,MONOOXYGENASES ,ENZYMES ,BIOCATALYSIS - Abstract
Cyclohexanone monooxygenase (CHMO) is a promising biocatalyst for industrial reactions owing to its broad substrate spectrum and excellent regio-, chemo-, and enantioselectivity. However, the low stability of many Baeyer-Villiger monooxygenases is an obstacle for their exploitation in industry. Characterization and crystal structure determination of a robust CHMO from Thermocrispum municipale is reported. The enzyme efficiently converts a variety of aliphatic, aromatic, and cyclic ketones, as well as prochiral sulfides. A compact substrate-binding cavity explains its preference for small rather than bulky substrates. Small-scale conversions with either purified enzyme or whole cells demonstrated the remarkable properties of this newly discovered CHMO. The exceptional solvent tolerance and thermostability make the enzyme very attractive for biotechnology. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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22. Expanding the Druggable Space of the LSD1/CoREST Epigenetic Target: New Potential Binding Regions for Drug-Like Molecules, Peptides, Protein Partners, and Chromatin.
- Author
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Robertson, James C., Hurley, Nate C., Tortorici, Marcello, Ciossani, Giuseppe, Borrello, Maria Teresa, Vellore, Nadeem A., Ganesan, A., Mattevi, Andrea, and Baron, Riccardo
- Subjects
LYSINE ,EPIGENETICS ,MOLECULAR dynamics ,BINDING sites ,MOLECULAR probes - Abstract
Lysine specific demethylase-1 (LSD1/KDM1A) in complex with its corepressor protein CoREST is a promising target for epigenetic drugs. No therapeutic that targets LSD1/CoREST, however, has been reported to date. Recently, extended molecular dynamics (MD) simulations indicated that LSD1/CoREST nanoscale clamp dynamics is regulated by substrate binding and highlighted key hinge points of this large-scale motion as well as the relevance of local residue dynamics. Prompted by the urgent need for new molecular probes and inhibitors to understand LSD1/CoREST interactions with small-molecules, peptides, protein partners, and chromatin, we undertake here a configurational ensemble approach to expand LSD1/CoREST druggability. The independent algorithms FTMap and SiteMap and our newly developed Druggable Site Visualizer (DSV) software tool were used to predict and inspect favorable binding sites. We find that the hinge points revealed by MD simulations at the SANT2/Tower interface, at the SWIRM/AOD interface, and at the AOD/Tower interface are new targets for the discovery of molecular probes to block association of LSD1/CoREST with chromatin or protein partners. A fourth region was also predicted from simulated configurational ensembles and was experimentally validated to have strong binding propensity. The observation that this prediction would be prevented when using only the X-ray structures available (including the X-ray structure bound to the same peptide) underscores the relevance of protein dynamics in protein interactions. A fifth region was highlighted corresponding to a small pocket on the AOD domain. This study sets the basis for future virtual screening campaigns targeting the five novel regions reported herein and for the design of LSD1/CoREST mutants to probe LSD1/CoREST binding with chromatin and various protein partners. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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23. Alternative Splicing of the Histone Demethylase LSD1/KDM1 Contributes to the Modulation of Neurite Morphogenesis in the Mammalian Nervous System.
- Author
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Zibetti, Cristina, Adamo, Antonio, Binda, Claudia, Forneris, Federico, Toffolo, Emanuela, Verpelli, Chiara, Ginelli, Enrico, Mattevi, Andrea, Sala, Carlo, and Battaglioli, Elena
- Subjects
CHROMATIN ,AMINO acids ,ENZYMES ,LYSINE ,AXONS - Abstract
A variety of chromatin remodeling complexes are thought to orchestrate transcriptional programs that lead neuronal precursors from earliest commitment to terminal differentiation. Here we show that mammalian neurons have a specialized chromatin remodeling enzyme arising from a neurospecific splice variant of LSD1/KDM1, histone lysine specific demethylase 1, whose demethylase activity on Lys4 of histone H3 has been related to gene repression. We found that alternative splicing of LSD1 transcript generates four full-length isoforms from combinatorial retention of two identified exons: the 4 aa exon E8a is internal to the amine oxidase domain, and its inclusion is restricted to the nervous system. Remarkably, the expression of LSD1 splice variants is dynamically regulated throughout cortical development, particularly during perinatal stages, with a progressive increase of LSD1 neurospecific isoforms over the ubiquitous ones. Notably, the same LSD1 splice dynamics can be fairly recapitulated in cultured cortical neurons. Functionally, LSD1 isoforms display in vitro a comparable demethylase activity, yet the inclusion of the sole exon E8a reduces LSD1 repressor activity on a reporter gene. Additional distinction among isoforms is supported by the knockdown of neurospecific variants in cortical neurons resulting in the inhibition of neurite maturation, whereas overexpression of the same variants enhances it. Instead, perturbation of LSD1 isoforms that are devoid of the neurospecific exon elicits no morphogenic effect. Collectively, results demonstrate that the arousal of neuronal LSD1 isoforms pacemakes early neurite morphogenesis, conferring a neurospecific function to LSD1 epigenetic activity. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
24. Flavoenzymes: diverse catalysts with recurrent features.
- Author
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Fraaije, Marco W. and Mattevi, Andrea
- Subjects
- *
FLAVOPROTEINS , *ENZYMES , *CHEMICAL structure - Abstract
Examines the three-dimensional structures of flavoenzymes and searches for recurrent features in their catalytic apparatus. Overview of the actions of flavoenzymes; Factors for the catalytic power of enzymes; Binding features of the isoalloxazine ring in flavin-dependent enzymes.
- Published
- 2000
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25. Crystallization and preliminary X-ray analysis of an alditol oxidase from Streptomyces coelicolor A3(2).
- Author
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Forneris, Federico, Rovida, Stefano, Heuts, Dominic P. H. M., Fraaije, Marco W., and Mattevi, Andrea
- Subjects
CRYSTALLIZATION ,OXIDASES ,STREPTOMYCES coelicolor ,CRYSTALS ,ENZYMES ,ALDOSE reductase - Abstract
Alditol oxidase is a 45 kDa enzyme containing a covalently bound FAD cofactor. This oxidase efficiently oxidizes a range of alditols to the corresponding aldoses. Owing to its substrate range and regioselectivity, this enzyme is an interesting candidate for biotechnological applications. Crystals of alditol oxidase from Streptomyces coelicolor A3(2) were obtained by the hanging-drop vapour-diffusion method and diffracted to 1.1 Å resolution. The crystals belong to space group C2, with unit-cell parameters a = 107, b = 68, c = 58 Å, β = 94°. Crystals of seleno-l-methionine-labelled alditol oxidase were obtained after seeding the crystallization drops with native microcrystals and showed a diffraction limit of 2.4 Å. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
26. Structural insights into the mechanism of amine oxidation by monoamine oxidases A and B
- Author
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Edmondson, Dale E., Binda, Claudia, and Mattevi, Andrea
- Subjects
- *
OXIDATION , *AMINES , *MONOAMINE oxidase , *ENZYMES - Abstract
Abstract: Due to their pharmacological importance in the oxidation of amine neurotransmitters, the membrane-bound flavoenzymes monoamine oxidase A and monoamine oxidase B have attracted numerous investigations and, as a result, two different mechanisms; the single electron transfer and the polar nucleophilic mechanisms, have been proposed to describe their catalytic mechanisms. This review compiles the recently available structural data on both enzymes with available mechanistic data as well as current NMR data on flavin systems to provide an integration of the approaches. These conclusions support the proposal that a polar nucleophilic mechanism for amine oxidation is the most consistent mechanistic scheme as compared with the single electron transfer mechanism. [Copyright &y& Elsevier]
- Published
- 2007
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27. Functional Role of the "Aromatic Cage" in Human Monoamine Oxidase B: Structures and Catalytic Properties of Tyr435 Mutant Proteins.
- Author
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Min Li, Binda, Claudia, Mattevi, Andrea, and Edmondson, Dale E.
- Subjects
- *
AMINO acids , *PHYSIOLOGICAL oxidation , *ENZYMES , *CATALYSIS , *GENETIC mutation , *PICHIA pastoris - Abstract
Current structural results of several flavin-dependent amine oxidizing enzymes including human monoamine oxidases A and B (MAO A and MAO B) show aromatic amino acid residues oriented approximately perpendicular to the flavin ring, suggesting a functional role in catalysis. In the case of human MAO B, two tyrosyl residues (Y398 and Y435) are found in the substrate binding site on the re face of the covalent flavin ring [Binda et al. (2002) J. Biol. Chem. 277, 23973-23976]. To probe the functional significance of this structure, Tyr435 in MAO B was mutated with the amino acids Phe, His, Leu, or Trp, the mutant proteins expressed in Pichia pastoris, and purified to homogeneity. Each mutant protein contains covalent FAD and exhibits a high level of catalytic functionality. No major alterations in active site structures are detected on comparison of their respective crystal structures with that of WT enzyme. The relative kcat/Km values for each mutant enzyme show Y435 > Y435F = Y435L = Y435H > Y435W. A similar behavior is also observed with the membrane-bound forms of MAO A and MAO B (MAO A Y444 mutant enzymes are found to be unstable on membrane extraction). p-Nitrobenzylamine is found to be a poor substrate while p-nitrophenethylamine is found to be a good substrate for all WT and mutant forms of MAO B. Analysis of these kinetic and structural data suggests the function of the "aromatic cage" in MAO to include a steric role in substrate binding and access to the flavin coenzyme and to increase the nucleophilicity of the substrate amine moiety. These results are consistent with a proposed polar nucleophilic mechanism for catalytic amine oxidation. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
28. Architecture of the NADPH oxidase family of enzymes.
- Author
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Heppner, David, Ogboo, Blessing, Grabovyy, Uriy, Maini, Aniket, Scouten, Scott, van der Vliet, Albert, and Mattevi, Andrea
- Subjects
- *
ENZYMES , *NADPH oxidase - Published
- 2022
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29. Finding the Switch: Turning a Baeyer-Villiger Monooxygenase into a NADPH Oxidase.
- Author
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Brondani, Patrícia B., Dudek, Hanna M., Martinoli, Christian, Mattevi, Andrea, and Fraaije, Marco W.
- Subjects
- *
ENZYMES , *CHEMICAL synthesis , *NADPH oxidase , *BAEYER-Villiger rearrangement , *MONOOXYGENASES , *FLAVINS , *INTERMEDIATES (Chemistry) - Abstract
By a targeted enzyme engineering approach, we were able to create an efficient NADPH oxidase from a monooxygenase. Intriguingly, replacement of only one specific single amino acid was sufficient for such a monooxygenase-to-oxidase switch--a complete transition in enzyme activity. Pre-steady-state kinetic analysis and elucidation of the crystal structure of the C65D PAMO mutant revealed that the mutation introduces small changes near the flavin cofactor, resulting in a rapid decay of the peroxyflavin intermediate. The engineered biocatalyst was shown to be a thermostable, solvent tolerant, and effective cofactor-regenerating biocatalyst. Therefore, it represents a valuable new biocatalytic tool. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
30. Molecular and Mechanistic Properties of the Membrane-Bound Mitochondrial Monoamine Oxidases.
- Author
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Edmondson, Dale E., Binda, Claudia, Jin Wang, Upadhyay, Anup K., and Mattevi, Andrea
- Subjects
- *
MONOAMINE oxidase , *MITOCHONDRIAL membranes , *PHARMACOLOGY , *ENZYMES , *NEUROPROTECTIVE agents - Abstract
The past decade has brought major advances in our knowledge of the structures and mechanisms of MAO A and MAO B, which are pharmacological targets for specific inhibitors. In both enzymes, crystallographic and biochemical data show their respective C-terminal transmembrane helices anchor the enzymes to the outer mitochondrial membrane. Pulsed EPR data show both enzymes are dimeric in their membrane-bound forms with agreement between distances measured in their crystalline forms. Distances measured between active site-directed spin-labels in membrane preparations show excellent agreement with those estimated from crystallographic data. Our knowledge of requirements for development of specific reversible MAO B inhibitors is in a fairly mature status. Less is known regarding the structural requirements for highly specific reversible MAO A inhibitors. In spite of their 70% level of sequence identity and similarities of Cα folds, the two enzymes exhibit significant functional and structural differences that can be exploited in the ultimate goal of the development of highly specific inhibitors. This review summarizes the current structural and mechanistic information available that can be utilized in the development of future highly specific neuroprotectants and cardioprotectants. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
31. Structural Analysis of the Catalytic Mechanism and Stereoselectivity in Streptomvces coelicolor Alditol Oxidase.
- Author
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Forneris, Federico, Heuts, Dominic P. H. M., Delvecchio, Manuela, Rovida, Stefano, Fraaije, Marco W., and Mattevi, Andrea
- Subjects
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STREPTOMYCES coelicolor , *FLAVINS , *OXIDATION , *HYDROXYL group , *ENZYMES - Abstract
Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex with both six-carbon (mannitol and sorbitol) and five-carbon substrates (xylitol). AldO shares the same folding topology of the members of the vanillyl-alcohol oxidase family of flavoenzymes and exhibits a covalently linked FAD which is located at the bottom of a funnel-shaped pocket that forms the active site. The high resolution of the three-dimensional structures highlights, a well-defined hydrogen-bonding network that tightly constrains the substrate in the productive conformation for catalysis. Substrate binding occurs through a lock-and-key mechanism and does not induce conformational changes with respect to the ligand-free protein. A network of charged residues is proposed to favor catalysis through stabilization of the deprotonated form of the substrate. A His side chain acts as back door that "pushes" the substrate- reactive carbon atom toward the N5-C4a locus of the flavin. Analysis of the three-dimensional structure reveals possible pathways for diffusion of molecular oxygen and a small cavity on the re side of the flavin that may host oxygen during FAD reoxidation. These features combined with the tight shape of the catalytic site provide insights into the mechanism of AldO-mediated regioselective oxidation reactions and its substrate specificity. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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32. A Highly Specific Mechanism of Histone H3-K4 Recognition by Histone Demethylase LSD1.
- Author
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Forneris, Federico, Binda, Claudia, Dall'Aglio, Annachiara, Fraaije, Marco W., Battaglioli, Elena, and Mattevi, Andrea
- Subjects
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HISTONES , *CHROMATIN , *METHYL groups , *ENZYMES , *BIOCHEMICAL research , *MOLECULAR biology - Abstract
Human lysine-specific demethylase (LSD1) is a chromatin-modifying enzyme that specifically removes methyl groups from mono- and dimethylated Lys4 of histone H3 (H3-K4). We used a combination of in vivo and in vitro experiments to characterize the substrate specificity and recognition by LSD1. Biochemical assays on histone peptides show that essentially all epigenetic modifications on the 21 N-terminal amino acids of histone H3 cause a significant reduction in enzymatic activity. Replacement of Lys4 with Arg greatly enhances binding affinity, and a histone peptide incorporating this mutation has a strong inhibitory power. Conversely, a peptide bearing a trimethylated Lys4 is only a weak inhibitor of the enzyme. Rapid kinetics measurements evidence that the enzyme is efficiently reoxidized by molecular oxygen with a second-order rate constant of 9.6 × 10³ M-1 s-1 and that the presence of the reaction product does not greatly influence the rate of flavin reoxidation. In vivo experiments provide a correlation between the in vitro inhibitory properties of the tested peptides and their ability of affecting endogenous LSD1 activity. Our results show that epigenetic modifications on histone H3 need to be removed before Lys4 demethylation can efficiently occur, The complex formed by LSD1 with histone deacetylases 1/2 may function as a ‘double-blade razor’ that first eliminates the acetyl groups from acetylated Lys residues and then removes the methyl group from Lys4. We suggest that after H3-K4 demethylation, LSD1 recruits the forthcoming chromatin remodelers leading to the introduction of gene repression marks. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
33. Role of the His57-Glu214 Ionic Couple Located in the Active Site of Mycobacterium tuberculosis FprA.
- Author
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Pennati, Andrea, Razeto, Adelia, De Rosa, Matteo, Pandini, Vittorio, Antonietta Vanoni, Maria, Mattevi, Andrea, Coda, Alessandro, Aliverti, Alessandro, and Zanetti, Giuliana
- Subjects
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FERREDOXIN-NADP reductase , *ENZYMES , *MYCOBACTERIUM tuberculosis , *CRYSTALLOGRAPHY , *CATALYSIS - Abstract
Mycobacterium tuberculosis FprA is a NADPH-ferredoxin reductase, functionally and structurally similar to the mammalian adrenodoxin reductase. It is presumably involved in supplying electrons to one or more of the pathogen's cytochrome P450s through reduced ferredoxins. It has been proposed on the basis of crystallographic data (Bossi, R. T., et al. (2002) Biochemistry 41, 8807-8818) that the highly conserved His57 and Glu214 whose side chains are H-bonded are involved in catalysis. Both residues were individually changed to nonionizable amino acyl residues through site-directed mutagenesis. Steady-state kinetics showed that the role of Glu214 in catalysis is negligible. On the contrary, the substitutions of His57 markedly impaired the catalytic efficiency of FprA for ferredoxin in the physiological reaction. Furthermore, they decreased the kcat/Km value for NADPH in the ferricyanide reduction. Rapid-reaction (stopped-flow) kinetic analysis of the isolated reductive half-reaction of wild- type and His57Gln forms of FprA with NADPH and NADH allowed a detailed description of the mechanism of enzyme-bound FAD reduction, with the identification of the intermediates involved. The His57Gln mutation caused a 6-fold decrease in the rate of hydride transfer from either NADPH or NADH to the enzyme-bound FAD cofactor. The 3D structure of FprA-H57Q, obtained at 1.8 Å resolution, explains the inefficient hydride transfer of the mutant in terms of a suboptimal geometry of the nicotinamide-isoalloxazine interaction in the active site. These data demonstrate the role of His57 in the correct binding of NADPH to FprA for the subsequent steps of the catalytic cycle to proceed at a high rate. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
34. The Active Conformation of Glutamate Synthase and its Binding to Ferredoxin
- Author
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van den Heuvel, Robert H.H., Svergun, Dmitri I., Petoukhov, Maxim V., Coda, Alessandro, Curti, Bruno, Ravasio, Sergio, Vanoni, Maria A., and Mattevi, Andrea
- Subjects
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AMMONIA , *ENZYMES - Abstract
Glutamate synthases (GltS) are crucial enzymes in ammonia assimilation in plants and bacteria, where they catalyze the formation of two molecules of l-glutamate from l-glutamine and 2-oxoglutarate. The plant-type ferredoxin-dependent GltS and the functionally homologous α subunit of the bacterial NADPH-dependent GltS are complex four-domain monomeric enzymes of 140–165 kDa belonging to the NH2-terminal nucleophile family of amidotransferases. The enzymes function through the channeling of ammonia from the N-terminal amidotransferase domain to the FMN-binding domain. Here, we report the X-ray structure of the Synechocystis ferredoxin-dependent GltS with the substrate 2-oxoglutarate and the covalent inhibitor 5-oxo-l-norleucine bound in their physically distinct active sites solved using a new crystal form. The covalent Cys1-5-oxo-l-norleucine adduct mimics the glutamyl-thioester intermediate formed during l-glutamine hydrolysis. Moreover, we determined a high resolution structure of the GltS:2-oxoglutarate complex. These structures represent the enzyme in the active conformation. By comparing these structures with that of GltS α subunit and of related enzymes we propose a mechanism for enzyme self-regulation and ammonia channeling between the active sites. X-ray small-angle scattering experiments were performed on solutions containing GltS and its physiological electron donor ferredoxin (Fd). Using the structure of GltS and the newly determined crystal structure of Synechocystis Fd, the scattering experiments clearly showed that GltS forms an equimolar (1:1) complex with Fd. A fundamental consequence of this result is that two Fd molecules bind consecutively to Fd-GltS to yield the reduced FMN cofactor during catalysis. [Copyright &y& Elsevier]
- Published
- 2003
- Full Text
- View/download PDF
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