The structure of L-amino acid oxidase reveals the substrate trajectory into an enantiomerically conserved active site

Pawelek, Peter D.; Cheah, Jaime H.; Coulombe, René; Macheroux, Peter; Ghisla, Sandro; Vrielink, Alice

doi:10.1093/emboj/19.16.4204

Cited by 234 publications

(258 citation statements)

References 41 publications

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“…The FAD-binding patterns of GO, DAAO, and MSOX share an overall similarity; in all these enzymes the FAD-binding domain contains the conserved Rossmann fold ␤␣␤ motif (␤1, ␣1, and ␤2) (33), which serves as a dinucleotide-binding motif (35). The central part of this consensus motif is the sequence GXGXXG (Gly 11 , Gly 13 , and Gly 16 of helix ␣1) with the N-terminal end of helix ␣1 pointing toward the FAD pyrophosphate moiety, as observed for other dinucleotide-binding proteins (35). The binding of FAD in GO is that typical of the GR 2 subfamily (34); the prosthetic group adopts an extended conformation with the isoalloxazine ring located at the interface between the FAD-binding domain and the substrate-binding domain, facing with its re-side the inner part of the substrate-binding cavity.…”

Section: Resultsmentioning

confidence: 99%

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Mörtl¹,

Diederichs

Welte

et al. 2004

Journal of Biological Chemistry

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Structure-function relationships of the flavoprotein glycine oxidase (GO), which was recently proposed as the first enzyme in the biosynthesis of thiamine in Bacillus subtilis, has been investigated by a combination of structural and functional studies. The structure of the GO-glycolate complex was determined at 1.8 Å, a resolution at which a sketch of the residues involved in FAD binding and in substrate interaction can be depicted. GO can be considered a member of the "amine oxidase" class of flavoproteins, such as D-amino acid oxidase and monomeric sarcosine oxidase. With the obtained model of GO the monomer-monomer interactions can be analyzed in detail, thus explaining the structural basis of the stable tetrameric oligomerization state of GO, which is unique for the GR 2 subfamily of flavooxidases. On the other hand, the three-dimensional structure of GO and the functional experiments do not provide the functional significance of such an oligomerization state; GO does not show an allosteric behavior. The results do not clarify the metabolic role of this enzyme in B. subtilis; the broad substrate specificity of GO cannot be correlated with the inferred function in thiamine biosynthesis, and the structure does not show how GO could interact with ThiS, the following enzyme in thiamine biosynthesis. However, they do let a general catabolic role of this enzyme on primary or secondary amines to be excluded because the expression of GO is not inducible by glycine, sarcosine, or D-alanine as carbon or nitrogen sources.Glycine oxidase (GO, 1 EC 1.4.3.19) is a flavoprotein consisting of four identical subunits (369 residues each) and containing one molecule of noncovalently bound FAD per 42-kDa protein molecule (1, 2). GO catalyzes a reaction similar to that of D-amino acid oxidase (DAAO, EC 1.4.3.3), a paradigm of the dehydrogenase-oxidase class of flavoproteins (for a recent review see Ref.3). Both enzymes catalyze the oxidative deamination of amino acids to yield the corresponding ␣-imino acids and, after hydrolysis, ␣-keto acids, ammonia (or primary amines), and hydrogen peroxide. Both enzymes show a high pK a for flavin N-3H ionization, do not bind covalently the FAD cofactor, and react readily with sulfite (1-3), but they differ in substrate specificity. In addition to neutral D-amino acids (e.g. D-alanine, D-proline, etc., which are also good substrates of DAAO), GO catalyzes the oxidation of primary and secondary amines (e.g. glycine, sarcosine, etc.) partially sharing the substrate specificity with monomeric sarcosine oxidase (MSOX, EC 1.5.3.1), an enzyme that catalyzes the oxidative demethylation of sarcosine to yield glycine, formaldehyde, and hydrogen peroxide (4). According to investigations of the substrate specificity and of the binding properties, the GO active site seems to preferentially accommodate amines of a small size, such as glycine and sarcosine (1, 2). GO follows a ternary complex sequential mechanism with glycine, sarcosine, and D-proline as substrates in which the rate of product dissociat...

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Section: Resultsmentioning

confidence: 99%

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Mörtl¹,

Diederichs

Welte

et al. 2004

Journal of Biological Chemistry

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“…In spite of the low sequence similarity, the topology of MAO, modeled from PAO, also closely resembles that of L-amino acid oxidase (42) with further similarities to D-amino acid oxidase (43) and p-hydroxybenzoate hydroxylase (44). Within this family of flavoproteins, most structural differences appear in the helical (interface) domain (42).…”

Section: Modeling Of Mao a And Mao B-mentioning

confidence: 99%

“…Within this family of flavoproteins, most structural differences appear in the helical (interface) domain (42).…”

Section: Modeling Of Mao a And Mao B-mentioning

confidence: 99%

Analysis of Conserved Active Site Residues in Monoamine Oxidase A and B and Their Three-dimensional Molecular Modeling

Geha¹,

Chen²,

Wouters³

et al. 2002

Journal of Biological Chemistry

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Monoamine oxidase (MAO) is a key enzyme responsible for the degradation of serotonin, norepinephrine, dopamine, and phenylethylamine. It is an outer membrane mitochondrial enzyme existing in two isoforms, A and B. We have recently generated 14 site-directed mutants of human MAO A and B, and we found that four key amino acids, Lys-305, Trp-397, Tyr-407, and Tyr-444, in MAO A and their corresponding amino acids in MAO B, Lys-296, Trp-388, Tyr-398, and Tyr-435, play important roles in MAO catalytic activity. Based on the polyamine oxidase three-dimensional crystal structure, it is suggested that Lys-305, Trp-397, and Tyr-407 in MAO A and Lys-296, Trp-388, and Tyr-398 in MAO B may be involved in the non-covalent binding to FAD. Tyr-407 and Tyr-444 in MAO A (Tyr-398 and Tyr-435 in MAO B) may form an aromatic sandwich that stabilizes the substrate binding. Asp-132 in MAO A (Asp-123 in MAO B) located at the entrance of the U-shaped substrate-binding site has no effect on MAO A nor MAO B catalytic activity. The similar impact of analogous mutants in MAO A and MAO B suggests that these amino acids have the same function in both isoenzymes. Three-dimensional modeling of MAO A and B using polyamine oxidase as template suggests that the overall tertiary structure and the active sites of MAO A and B may be similar.Monoamine oxidase (MAO, 1 EC 1.4.3.4; amine:oxygen oxidoreductase (deaminating, flavin-containing)) is a flavoprotein located at the outer membranes of mitochondria in neuronal, glial, and other cells. It catalyzes the oxidative deamination of monoamine neurotransmitters such as serotonin, norepinephrine, and dopamine and appears to play important roles in several psychiatric and neurological disorders (for review see Refs. 1 and 2). In addition, it is also responsible for the biotransformation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine into 1-methyl-4-phenylpyridinium, a Parkinsonian producing neurotoxin (3-5). Recently, it has been shown that MAO may contribute to the apoptotic process because inhibition of MAO activity suppressed cell death (6).MAO exists in two forms, namely MAO A and MAO B. MAO A preferentially oxidizes serotonin (5-hydroxytryptamine) and is irreversibly inhibited by low concentrations of clorgyline (7). MAO B preferentially oxidizes phenylethylamine (PEA) and benzylamine, and it is irreversibly inactivated by low concentrations of pargyline and deprenyl (8). Dopamine, tyramine, and tryptamine are common substrates for both MAOs. MAO A and B consist of 527 and 520 amino acids, respectively, and have a 70% identity (9). Each isoenzyme has a FAD covalently linked to a cysteine residue, Cys-406 in MAO A and Cys-397 in MAO B, through an 8␣-(cysteinyl)-riboflavin (10 -13). They exhibit identical exon-intron organization, and they are probably derived from the duplication of a common ancestral gene (14 Recently, the x-ray crystal structure of a related enzyme namely polyamine oxidase (PAO) has been obtained (25). PAO catalyzes the oxidation of the secondary amino group of polyamines, such as sp...

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“…It constitutes 1-9% of the total venom protein and is responsible for the light yellowish color of the venom and catalyzes the stereospecific de-amination of an L-amino acid substrate to an alpha-keto acid along with the production of ammonia and hydrogen peroxide. It has been found that LAAO from snake venom can induce apoptosis in mammalian endothelial cells possible due to the production of high concentration of hydrogen peroxide (Pawelek et al, 2000).…”

Section: L-amino Acid Oxidase (Laao)mentioning

confidence: 99%

“…LAAOs are dimeric flavoprotein that contains a non-covalently bound FAD as a co-factor (Pawelek et al, 2000). LAAOs isolated from Ophiophagus hannah venom decreases thymidine uptake in murine melanoma, fibrosarcoma, colorectal cancer and Chinese hamster ovary cell line that also showed reduction in cellular proliferation (Cura et al, 2002).…”

Section: Anticancer Activity Of Snake Venommentioning

confidence: 99%

Snake Venom: A Potent Anticancer Agent

Jain

Kumar²

2012

Asian Pacific Journal of Cancer Prevention

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Since cancer is one of the leading causes of death worldwide, and there is an urgent need to find better treatment. In recent years remarkable progress has been made towards the understanding of proposed hallmarks of cancer development and treatment. Treatment modalities comprise radiation therapy, surgery, chemotherapy, immunotherapy and hormonal therapy. Currently, the use of chemotherapeutics remains the predominant option for clinical control. However, one of the major problems with successful cancer therapy using chemotherapeutics is that patients often do not respond or eventually develop resistance after initial treatment. This has led to the increased use of anticancer drugs developed from natural resources. The biodiversity of venoms and toxins makes them a unique source from which novel therapeutics may be developed. In this review, the anticancer potential of snake venom is discussed. Some of the included molecules are under clinical trial and may find application for anticancer drug development in the near future.

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The structure of L-amino acid oxidase reveals the substrate trajectory into an enantiomerically conserved active site

Cited by 234 publications

References 41 publications

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Analysis of Conserved Active Site Residues in Monoamine Oxidase A and B and Their Three-dimensional Molecular Modeling

Snake Venom: A Potent Anticancer Agent

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