HPLC and NMR Investigation of the Serum Amine Oxidase Catalyzed Oxidation of Polyamines.

Bock, K. D.; Jensen, Arne L.; Weidlein, Johann; Schnöckel, Hansgeorg; Paulsen, Gudrun B.; Nielsen, Ruby I.; Olsen, Carl Erik; Pedersen, Christian; Stidsen, Carsten E.

doi:10.3891/acta.chem.scand.48-0052

Cited by 45 publications

(56 citation statements)

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“…The factors favoring its elimination from the oxidation products of spermine and spermidine have not been identified. The direct oxidation of spermine to spermidine and 3-aminopropanal by BSAO was reported by Houen et al (26).…”

Section: Treatments and Cell Survival Experimentsmentioning

confidence: 66%

Sensitization of human colon adenocarcinoma cells (LoVo) to reactive oxygen species by a lysosomotropic compound

Agostinelli

Vedova²,

Belli³

et al. 2006

Int J Oncol

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Abstract.The in situ formation of cytotoxic metabolites by an enzyme-catalyzed reaction is a recent approach in cancer therapy. The present results show that multidrug-resistant human colon adenocarcinoma cells (LoVo) are significantly more sensitive than corresponding wild-type cells to hydrogen peroxide and aldehydes, the products of bovine serum amine oxidase (BSAO)-catalyzed oxidation of spermine. Pretreatment of the cells with N 1 ,N 4 -bis(2,3-butadienyl)-1,4-butanediamine (MDL 72527), a lysosomotropic compound, sensitized both cell lines to the subsequent exposure to spermine metabolites, as was evident from the decrease of cell survival by a log unit. The sensitizing effect was greater in the case of the multidrug-resistant cell line, an aspect of particular importance with respect to potential therapeutic applications of the method, since conventional cancer therapy suffers from the development of drug resistance. Cell viability was determined using a clonogenic assay. MDL 72527 (at 300 μM) produced numerous cytoplasmic vacuoles, presumably of lysosomal origin, after 6-h exposure, which decreased in size and number (in the presence of the drug) by 24 h and had almost disappeared completely at 48 h. Mitochondrial damage, as observed by transmission electron microscopy, seemed to correlate better with the cytotoxic effects of the treatment than the formation of vacuoles. We suggest that the release of lysosomal enzymes into the cytosol by MDL 72527 is the main reason for its sensitizing effect. It is known that lysosomotropic compounds, which release lysosomal enzymes, produce oxidative stress and apoptosis.

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Section: Treatments and Cell Survival Experimentsmentioning

confidence: 66%

Sensitization of human colon adenocarcinoma cells (LoVo) to reactive oxygen species by a lysosomotropic compound

Agostinelli

Vedova²,

Belli³

et al. 2006

Int J Oncol

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“…This hydrolysis would produce 3-aminopropanal, which can spontaneously convert to acrolein (18). However, the 2,4-dinitrophenylhydrazones of either 3-aminopropanal or acrolein were not detected in the HPLC analyses of the enzyme reaction solutions (Fig.…”

Section: Heterologous Production Of Mpao By E Coli and Characterizatmentioning

confidence: 99%

“…Thus, PAO has been proposed to play a role in triggering and/or participating in the progression of apoptosis (13)(14)(15)(16)(17). Additionally, another product of the oxidation of the N 1 -acetylated polyamines, 3-acetamidopropanal, may be enzymatically deacetylated to yield cytotoxic 3-aminopropanal (18) that is thought to contribute, either alone or in concert with H 2 O 2 , to tissue damage following traumatic injury (19 -21).…”

mentioning

confidence: 99%

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Cloning, Sequencing, and Heterologous Expression of the Murine Peroxisomal Flavoprotein, N1-Acetylated Polyamine Oxidase

Wu¹,

Yankovskaya²,

McIntire³

2003

Journal of Biological Chemistry

113

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The aminoacyl sequences of three regions of pure bovine N 1 -acetylated polyamine oxidase (PAO) were obtained and used to search GenBank TM . This led to the cloning and sequencing of a complete coding cDNA for murine PAO (mPAO) and the 5-truncated coding region of the bovine pao (bpao) gene. A search of GenBank TM indicated that mpao maps to murine chromosome 7 as seven exons. The translated amino acid sequences of mpao and bpao have a -Pro-Arg-Leu peroxisomal targeting signal at the extreme C termini. A ␤-␣-␤ FAD-binding motif is present in the N-terminal portion of mPAO. This and several other regions of mPAO and bPAO are highly similar to corresponding sections of other flavoprotein amine oxidases, although the overall identity of aligned sequences indicates that PAO represents a new subfamily of flavoproteins. A fragment of mpao was used as a probe to establish the relative transcription levels of this gene in various mature murine tissues and murine embryonic and breast tissues at different developmental stages. An Escherichia coli expression system has been developed for manufacturing mPAO at a reasonable level. The mPAO so produced was purified to homogeneity and characterized. It was demonstrated definitively that PAO oxidizes N 1 -acetylspermine to spermidine and 3-acetamidopropanal and that it also oxidizes N 1 -acetylspermidine to putrescine and 3-acetamidopropanal. Thus, this is the classical polyamine oxidase (EC 1.5.3.11) that is defined as the enzyme that oxidizes these N 1 -acetylated polyamines on the exo-side of their N 4 -amino groups. This enzyme is distinguishable from the plant polyamine oxidase that oxidizes spermine on the endo-side of the N 4 -nitrogen. It differs also from mammalian spermine oxidase that oxidizes spermine (but not N 1 -acetylspermine or N 1 -acetylspermidine) at the exo-carbon of its N 4 -amino group. This report provides details of the biochemical, spectral, oxidationreduction, and steady-state kinetic properties of pure mPAO. Spermine (SPM)1 and spermidine (SPD) are important polyamines required for numerous fundamentally important cellular processes including wound healing, tissue growth, tissue differentiation, and tumor growth (1-9). In mammalian cells, polyamine pool homeostasis is maintained by a balance of enzymatic and transport processes as follows.

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“…3-Aminopropanal appears to be the main metabolite of polyamine catabolism mediated by FAD-dependent enzymes, which can dissociate into acrolein and ammonia [53]. There has been some dispute as to whether spermine is metabolized by copper-dependent plasma amine oxidase at the primary amino termini or at the secondary amine sites [54][55][56]. Both pathways would yield spermidine, acrolein, and 3-aminopropanal via an intermediate imine species followed by hydrolysis and addition/elimination reactions.…”

Section: Polyaminesmentioning

confidence: 99%

Acrolein: Sources, metabolism, and biomolecular interactions relevant to human health and disease

Stevens

Maier

2008

Molecular Nutrition Food Res

632

634

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Acrolein (2-propenal) is ubiquitously present in (cooked) foods and in the environment. It is formed from carbohydrates, vegetable oils and animal fats, amino acids during heating of foods, and by combustion of petroleum fuels and biodiesel. Chemical reactions responsible for release of acrolein include heat-induced dehydration of glycerol, retro-aldol cleavage of dehydrated carbohydrates, lipid peroxidation of polyunsaturated fatty acids, and Strecker degradation of methionine and threonine. Smoking of tobacco products equals or exceeds the total human exposure to acrolein from all other sources. The main endogenous sources of acrolein are myeloperoxidase-mediated degradation of threonine and amine oxidase-mediated degradation of spermine and spermidine, which may constitute a significant source of acrolein in situations of oxidative stress and inflammation. Acrolein is metabolized by conjugation with glutathione and excreted in the urine as mercapturic acid metabolites. Acrolein forms Michael adducts with ascorbic acid in vitro, but the biological relevance of this reaction is not clear. The biological effects of acrolein are a consequence of its reactivity towards biological nucleophiles such as guanine in DNA and cysteine, lysine, histidine, and arginine residues in critical regions of nuclear factors, proteases, and other proteins. Acrolein adduction disrupts the function of these biomacromolecules which may result in mutations, altered gene transcription, and modulation of apoptosis.

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HPLC and NMR Investigation of the Serum Amine Oxidase Catalyzed Oxidation of Polyamines.

Cited by 45 publications

References 0 publications

Sensitization of human colon adenocarcinoma cells (LoVo) to reactive oxygen species by a lysosomotropic compound

Sensitization of human colon adenocarcinoma cells (LoVo) to reactive oxygen species by a lysosomotropic compound

Cloning, Sequencing, and Heterologous Expression of the Murine Peroxisomal Flavoprotein, N1-Acetylated Polyamine Oxidase

Acrolein: Sources, metabolism, and biomolecular interactions relevant to human health and disease

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