Publications

• Mammalian plasma and tissue-bound semicarbazide-sensitive amine oxidases: biochemical, pharmacological and toxicological aspects.

  Lyles G.A.

  Mammalian plasma and tissues contain various soluble and membrane-bound enzymes which metabolize the synthetic amine benzylamine particularly well. The sensitivity of these enzymes to inhibition by semicarbazide and related compounds suggests that they contain a cofactor with a reactive carbonyl g ... >> More

  Mammalian plasma and tissues contain various soluble and membrane-bound enzymes which metabolize the synthetic amine benzylamine particularly well. The sensitivity of these enzymes to inhibition by semicarbazide and related compounds suggests that they contain a cofactor with a reactive carbonyl group, which has been proposed to be either pyridoxal phosphate, pyrroloquinoline quinone or (more recently) 6-hydroxydopa. It is not yet clear if all of these semicarbazide-sensitive amine oxidases (SSAOs) are copper-dependent enzymes. A variety of compounds have now been identified as relatively selective inhibitors to distinguish the SSAOs from other amine oxidases, in order to investigate the properties of SSAOs and their potential role in biogenic and xenobiotic amine metabolism in vivo. While plasma SSAO is soluble, most tissue SSAOs appear to be membrane-bound, probably plasmalemmal enzymes, which may be capable of metabolizing extracellular amines. Vascular (and non-vascular) smooth muscle cells have particularly high SSAO activity, although recently the enzyme has been found in other cell types (e.g. adipocytes, chondrocytes, odontoblasts) implying a functional importance not restricted solely to smooth muscle. The substrate specificity of plasma and tissue SSAOs shows considerable species-related variations. For example, while some endogenously-occurring aromatic amines such as tyramine and tryptamine are metabolized well by SSAO in homogenates of rat blood vessels, and also in vitro inhibition of SSAO can potentiate vasoconstrictor actions of these amines in rat vascular preparations, these amines are poor substrates for human SSAO, thus complicating attempts to generalize possible physiological roles for these enzymes. Vascular SSAO can metabolize the xenobiotic aliphatic amine, allylamine, to the cytotoxic aldehyde acrolein and this has been linked to the ability of allylamine administration to produce cardiovascular lesions in experimental animals, sometimes mimicking features of atherosclerotic disease. Recent studies showing that the endogenously-occurring aliphatic amines methylamine and aminoacetone are metabolized in vitro to formaldehyde and methylglyoxal, respectively, by SSAO in some animal (including human) tissues, suggest the possibility that toxicological consequences upon cellular function could result if such conversions occur in vivo. << Less

  Int J Biochem Cell Biol 28:259-274(1996) [PubMed] [EuropePMC]

• Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications.

  Airenne T.T., Nymalm Y., Kidron H., Smith D.J., Pihlavisto M., Salmi M., Jalkanen S., Johnson M.S., Salminen T.A.

  The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs i ... >> More

  The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. << Less

  Protein Sci. 14:1964-1974(2005) [PubMed] [EuropePMC]

• Cloning and molecular analysis of the pea seedling copper amine oxidase.

  Tipping A.J., McPherson M.J.

  A pea seedling amine oxidase cDNA has been isolated and sequenced. A single long open reading frame has amino acid sequences corresponding to those determined from active site peptide (Janes, S.M., Palcic, M.M., Scaman, C.H., Smith, A.J., Brown, D.E., Dooley, D.M., Mure, M., and Klinman, J.P. (199 ... >> More

  A pea seedling amine oxidase cDNA has been isolated and sequenced. A single long open reading frame has amino acid sequences corresponding to those determined from active site peptide (Janes, S.M., Palcic, M.M., Scaman, C.H., Smith, A.J., Brown, D.E., Dooley, D.M., Mure, M., and Klinman, J.P. (1992) Biochemistry 31, 12147-12154) and N-terminal sequencing experiments. The latter reveals the protein to have a 25-amino acid leader sequence with characteristics of a secretion signal peptide, as expected for this extracellular enzyme. Comparisons of the amino acid sequence of the mature pea enzyme (649 amino acids) with that of the mature lentil enzyme (569 amino acids; Rossi, A., Petruzzelli, R., and Finazzi-Agrò, A. (1992) FEBS Lett. 301, 253-257) reveal important and unexpected differences particularly with regard to protein length. Sequencing of part of the lentil gene identified several frameshift differences within the coding region resulting in a mature lentil protein of exactly the same length, 649 amino acids, as the pea enzyme. Multiple alignments of 10 copper amine oxidase sequences reveal 33 completely conserved residues of which 10 are found within 41 aligned residues at the C-terminal tails, the region missing from the original lentil sequence. One of only four conserved histidines is found in this region and may represent the third ligand to the copper. The pea enzyme contains around 3-4% carbohydrate as judged by deglycosylation experiments. We have also demonstrated by hybridization analysis that copper amine oxidase genes are present in a range of mono- and dicotyledonous plants. << Less

  J. Biol. Chem. 270:16939-16946(1995) [PubMed] [EuropePMC]

• Reaffirmation that metabolism of polyamines by bovine plasma amine oxidase occurs strictly at the primary amino termini.

  Lee Y., Sayre L.M.

  Oxidation of the biologically important polyamines spermine and spermidine by plasma amine oxidase (PAO) was specified many years ago to occur at the terminal primary rather than internal secondary amine positions. However, the finding of sequential enzymatic conversion of spermine to spermidine a ... >> More

  Oxidation of the biologically important polyamines spermine and spermidine by plasma amine oxidase (PAO) was specified many years ago to occur at the terminal primary rather than internal secondary amine positions. However, the finding of sequential enzymatic conversion of spermine to spermidine and then to putrescine (1, 4-butanediamine) is superficially suggestive of metabolism at the secondary amine positions, and a recent publication (Houen, G., Bock, K., and Jensen, A. L. (1994) Acta Chem. Scand. 48, 52-60) claimed that the original interpretation of preferential "terminal" deamination does not stand up to scrutiny with modern methods of analysis. We herein demonstrate that the findings cited in support of secondary amine deamination can arise artifactually from spontaneous elimination/addition reactions following initial metabolism at the terminal positions of 3-(aminopropyl)amines. We further find no evidence for the ability of PAO to metabolize the secondary amine position in homospermidine, which is devoid of such complicating side reactions. Our results support the original claimed specificity of PAO for the primary amino termini of polyamines, all of which are consistent with the general finding that the quinone-dependent copper amine oxidases specifically metabolize primary amines. << Less

  J Biol Chem 273:19490-19494(1998) [PubMed] [EuropePMC]

• Tissue activity and cellular localization of human semicarbazide-sensitive amine oxidase.

  Andres N., Lizcano J.M., Rodriguez M.J., Romera M., Unzeta M., Mahy N.

  Semicarbazide-sensitive amine oxidase (SSAO), widely distributed in highly vascularized mammalian tissues, metabolizes endogenous and xenobiotic aromatic and aliphatic monoamines. To assess whether its physiological role in humans is restricted to oxidation, we used an immunohistochemical approach ... >> More

  Semicarbazide-sensitive amine oxidase (SSAO), widely distributed in highly vascularized mammalian tissues, metabolizes endogenous and xenobiotic aromatic and aliphatic monoamines. To assess whether its physiological role in humans is restricted to oxidation, we used an immunohistochemical approach to examine the cellular localization of SSAO in human peripheral tissues (adrenal gland, duodenum, heart, kidney, lung, liver, pancreas, spleen, thyroid gland, and blood vessels) and also analyzed its subcellular localization. The results are in agreement with the specific activities also determined in the same samples and are discussed with reference to the tissue distribution of monoamine oxidase A and B. Together with the oxidative deamination of monoamines, SSAO cellular localization indicates that, in most human peripheral tissues, it might participate in the regulation of physiological processes via H(2)O(2) generation. (J Histochem Cytochem 49:209-217, 2001) << Less

  J Histochem Cytochem 49:209-217(2001) [PubMed] [EuropePMC]

• Probing the catalytic mechanism of Escherichia coli amine oxidase using mutational variants and a reversible inhibitor as a substrate analogue.

  Saysell C.G., Tambyrajah W.S., Murray J.M., Wilmot C.M., Phillips S.E., McPherson M.J., Knowles P.F.

  Copper amine oxidases are homodimeric enzymes containing one Cu(2+) ion and one 2,4,5-trihydroxyphenylalanine quinone (TPQ) per monomer. Previous studies with the copper amine oxidase from Escherichia coli (ECAO) have elucidated the structure of the active site and established the importance in ca ... >> More

  Copper amine oxidases are homodimeric enzymes containing one Cu(2+) ion and one 2,4,5-trihydroxyphenylalanine quinone (TPQ) per monomer. Previous studies with the copper amine oxidase from Escherichia coli (ECAO) have elucidated the structure of the active site and established the importance in catalysis of an active-site base, Asp-383. To explore the early interactions of substrate with enzyme, we have used tranylcypromine (TCP), a fully reversible competitive inhibitor, with wild-type ECAO and with the active-site base variants D383E and D383N. The formation of an adduct, analogous to the substrate Schiff base, between TCP and the TPQ cofactor in the active site of wild-type ECAO and in the D383E and D383N variants has been investigated over the pH range 5.5-9.4. For the wild-type enzyme, the plot of the binding constant for adduct formation (K(b)) against pH is bell-shaped, indicating two pK(a)s of 5.8 and approximately 8, consistent with the preferred reaction partners being the unprotonated active-site base and the protonated TCP. For the D383N variant, the reaction pathway involving unprotonated base and protonated TCP cannot occur, and binding must follow a less favoured pathway with unprotonated TCP as reactant. Surprisingly, for the D383E variant, the K(b) versus pH behaviour is qualitatively similar to that of D383N, supporting a reaction pathway involving unprotonated TCP. The TCP binding data are consistent with substrate binding data for the wild type and the D383E variant using steady-state kinetics. The results provide strong support for a protonated amine being the preferred substrate for the wild-type enzyme, and emphasize the importance of the active-site base, Asp-383, in the primary binding event. << Less

  Biochem J 365:809-816(2002) [PubMed] [EuropePMC]

• Mammalian Cu-containing amine oxidases (CAOs): new methods of analysis, structural relationships, and possible functions.

  Houen G.

  This thesis describes new and original experimental results on Cu-dependent amine oxidases (CAOs), which show that these enzymes can be conveniently and specifically detected in situ using a peroxidase-coupled activity staining method with 4-Cl-1-naphtole as hydrogen donor substrate. Even more sen ... >> More

  This thesis describes new and original experimental results on Cu-dependent amine oxidases (CAOs), which show that these enzymes can be conveniently and specifically detected in situ using a peroxidase-coupled activity staining method with 4-Cl-1-naphtole as hydrogen donor substrate. Even more sensitive in situ detection can be achieved using a chemiluminescence-based coupled peroxidase assay which was applied to show that human placenta CAO activity is confined to maternal vessels. A general purification scheme for CAOs is described, and applied to purification of different CAOs. Peptide maps and immunological crossreactivity studies with monoclonal antibodies raised against the purified enzymes showed that they were closely related. Amino acid sequence data for the bovine serum CAO showed that they form a separate group (E.C. 1.4.3.6) with no homology to other enzymes. A cDNA sequence was obtained on the basis of the amino acid sequence data, and this was found to encode a bovine lung CAO, related to bovine serum CAO. The genes for bovine lung and bovine serum CAO are characterized, and Southern blotting analysis of bovine chromosomal DNA shows the existence of a least one more bovine CAO. The purification of human neutrophil CAO is attempted, but it is described how lactoferrin, a protein with many properties in common with CAOs, and with a low degree of sequence identity can account for many observations on human neutrophil CAO. The products of bovine serum CAO oxidation of polyamines are characterised, and 3-aminopropanal is found to be the principal aminoaldehyde produced. Finally, a polyamine-stimulated binding of human placenta CAO to single-stranded DNA is described, and it is reported that the DNA-bound CAO is enzymically active and that the oxidation of DNA-bound polyamines leads to degradation of DNA. In addition to the experimental results, the properties of polyamines and Cu-dependent amine oxidases are reviewed. The polyamines spermidine and spermine interact specifically with nucleic acids and several other molecules. They are synthesised from putrescine, which is a key regulatory molecule formed from ornithine by ornithine decarboxylase, a highly inducible and regulated enzyme. The polyamines can be converted to putrescine by CAOs or spermidine/spermine acetyltransferase and polyamine oxidase. Putrescine is degraded by CAOs, which are also involved in degradation of histamine, a mediator of inflammatory processes. CAOs catalyse the general reaction: R1CH2NHR2 + O2 + H2O-->R1CHO + R2NH2 + H2O2 and in addition to the catabolism of putrescine and histamine CAOs are involved in regulation of growth and apoptosis by to the generation of aminoaldehydes and hydrogen peroxide which have growth inhibitory properties. Several homologous CAOs have been purified and characterized and they form a family with two subgroups. They are homodimers with a relative molecular weight of 180,000 and contain Cu2+ and a modified tyrosine, topaquinone, in the active site. CAOs are present in most tissues with highest amounts in intestine, kidneys, liver and placenta, but the cellular distributions and functions of CAOs are still poorly described, partly due to the use of many different assays and partly due to a broad substrate specificity of the enzymes. However, polyamines and CAOs seem to form a universal system contributing to regulation of growth, differentiation, and apoptosis. << Less

  APMIS Suppl 96:1-46(1999) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Semicarbazide-sensitive amine oxidases: enzymes with quite a lot to do.

  O'Sullivan J., Unzeta M., Healy J., O'Sullivan M.I., Davey G., Tipton K.F.

  The semicarbazide-sensitive amine oxidases (SSAO) (EC 1.4.3.6) were believed to be detoxifying enzymes, primarily involved in the oxidative deamination of endogenous amines, such as methylamine and aminoacetone, together with some xenobiotic amines. However, it appears that the reaction products m ... >> More

  The semicarbazide-sensitive amine oxidases (SSAO) (EC 1.4.3.6) were believed to be detoxifying enzymes, primarily involved in the oxidative deamination of endogenous amines, such as methylamine and aminoacetone, together with some xenobiotic amines. However, it appears that the reaction products may have important signalling functions in the regulation of cell development and glucose homeostasis. Furthermore, enzyme, from some sources, behaves as a cellular adhesion protein under inflammatory and it may also be involved in lipid transport. This review considers what is known about the activities and potential functions of this hardworking protein. << Less

  Neurotoxicology 25:303-315(2004) [PubMed] [EuropePMC]

• Crystal structures of the copper-containing amine oxidase from Arthrobacter globiformis in the holo and apo forms: implications for the biogenesis of topaquinone.

  Wilce M.C., Dooley D.M., Freeman H.C., Guss J.M., Matsunami H., McIntire W.S., Ruggiero C.E., Tanizawa K., Yamaguchi H.

  The crystal structures of the copper enzyme phenylethylamine oxidase from the Gram-positive bacterium Arthrobacter globiformis (AGAO) have been determined and refined for three forms of the enzyme: the holoenzyme in its active form (at 2.2 A resolution), the holoenzyme in an inactive form (at 2.8 ... >> More

  The crystal structures of the copper enzyme phenylethylamine oxidase from the Gram-positive bacterium Arthrobacter globiformis (AGAO) have been determined and refined for three forms of the enzyme: the holoenzyme in its active form (at 2.2 A resolution), the holoenzyme in an inactive form (at 2.8 A resolution), and the apoenzyme (at 2.2 A resolution). The holoenzyme has a topaquinone (TPQ) cofactor formed from the apoenzyme by the post-translational modification of a tyrosine residue in the presence of Cu2+. Significant differences between the three forms of AGAO are limited to the active site. The polypeptide fold is closely similar to those of the amine oxidases from Escherichia coli [Parsons, M. R., et al. (1995) Structure 3, 1171-1184] and pea seedlings [Kumar, V., et al. (1996) Structure 4, 943-955]. In the active form of holo-AGAO, the active-site Cu atom is coordinated by three His residues and two water molecules in an approximately square-pyramidal arrangement. In the inactive form, the Cu atom is coordinated by the same three His residues and by the phenolic oxygen of the TPQ, the geometry being quasi-trigonal-pyramidal. There is evidence of disorder in the crystals of both forms of holo-AGAO. As a result, only the position of the aromatic group of the TPQ cofactor, but not its orientation about the Cbeta-Cgamma bond, is determined unequivocally. In apo-AGAO, electron density consistent with an unmodified Tyr occurs at a position close to that of the TPQ in the inactive holo-AGAO. This observation has implications for the biogenesis of TPQ. Two features which have not been described previously in amine oxidase structures are a channel from the molecular surface to the active site and a solvent-filled cavity at the major interface between the two subunits of the dimer. << Less

  Biochemistry 36:16116-16133(1997) [PubMed] [EuropePMC]

• Microbial oxidation of amines. Distribution, purification and properties of two primary-amine oxidases from the yeast Candida boidinii grown on amines as sole nitrogen source.

  Haywood G.W., Large P.J.

  1. The yeast Candida boidinii was grown on glucose as carbon source with a range of amines and amino acids as nitrogen sources. Cells grown on amines contained elevated activities of catalase. If the amines contained N-methyl groups, formaldehyde dehydrogenase, formate dehydrogenase and S-formylgl ... >> More

  1. The yeast Candida boidinii was grown on glucose as carbon source with a range of amines and amino acids as nitrogen sources. Cells grown on amines contained elevated activities of catalase. If the amines contained N-methyl groups, formaldehyde dehydrogenase, formate dehydrogenase and S-formylglutathione hydrolase were also elevated in activity compared with cells grown on (NH(4))(2)SO(4). 2. Cells grown on all the amines tested, but not those grown on urea or amino acids, contained an oxidase attacking primary amines, which is referred to as methylamine oxidase. In addition, cells grown on some amines contained a second amine oxidase, which is referred to as benzylamine oxidase. 3. Both amine oxidases were purified to near homogeneity. 4. Benzylamine oxidase was considerably more stable at 45 and 50 degrees C than was methylamine oxidase. 5. Both enzymes had a pH optimum in the region of 7.0, and had a considerable number of substrates in common. There were, however, significant differences in the substrate specificity of the two enzymes. The ratio V/K(app.) (m) increased with increasing n-alkyl carbon chain length for benzylamine oxidase, but decreased for methylamine oxidase. 6. Both enzymes showed similar sensitivity to carbonyl-group reagents, copper-chelating agents and other typical ;diamine oxidase inhibitors'. 7. The stoicheiometry for the reaction catalysed by each enzyme was established. 8. The kinetics of methylamine oxidase were examined by varying the methylamine and oxygen concentrations in turn. A non-Ping Pong kinetic pattern with intersecting double-reciprocal plots was obtained, giving K(m) values of 10mum for O(2) and 198mum for methylamine. The significance of this unusual kinetic behaviour is discussed. Similar experiments were not possible with the benzylamine oxidase, because it seemed to have an even lower K(m) for O(2). 9. Both enzymes had similar subunit M(r) values of about 80000, but the benzylamine oxidase behaved as if it were usually a dimer, M(r) 136000, which under certain conditions aggregated to a tetramer, M(r) 288000. Methylamine oxidase was mainly in the form of an octamer, M(r) 510000, which gave rise quite readily to dimers of M(r) 150000, and on gel filtration behaved as if the M(r) was 286000. << Less

  Biochem J 199:187-201(1981) [PubMed] [EuropePMC]