Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin

Vujcic, Slavoljub; Diegelman, Paula; Bacchi, Cyrus J.; Kramer, Debora L.; Porter, Carl W.

doi:10.1042/bj20020720

Cited by 197 publications

(190 citation statements)

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“…In fact, the targeted cells appeared to convert spermine to spermidine much more efficiently than did their wild-type counterparts (19). The conversion of spermine to spermidine in the absence of SSAT activity is obviously attributable to a recently discovered oxidase, which, when first cloned, was thought to be PAO (20), but was later identified as a novel flavin-containing spermine oxidase (21). Unlike PAO, SMO strongly prefers spermine to its acetylated derivatives.…”

Section: Discussionmentioning

confidence: 99%

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Metabolic Stability of α-Methylated Polyamine Derivatives and Their Use as Substitutes for the Natural Polyamines

Järvinen

Grigorenko

Khomutov

et al. 2005

Journal of Biological Chemistry

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Metabolically stable polyamine derivatives may serve as useful surrogates for the natural polyamines in studies aimed to elucidate the functions of individual polyamines. Here we studied the metabolic stability of ␣-methylspermidine, ␣-methylspermine, and bis-␣-methylspermine, which all have been reported to fulfill many of the putative physiological functions of the natural polyamines. In vivo studies were performed with the transgenic rats overexpressing spermidine/spermine N 1 -acetyltransferase. ␣-Methylspermidine effectively accumulated in the liver and did not appear to undergo any further metabolism. On the other hand, ␣-methylspermine was readily converted to ␣-methylspermidine and spermidine; similarly, bis-␣-methylspermine was converted to ␣-methylspermidine to some extent, both conversions being inhibited by the polyamine oxidase inhibitor N 1 ,N 2 -bis(2,3-butadienyl)-1,4-butanediamine. Furthermore, we used recombinant polyamine oxidase, spermidine/spermine N 1 -acetyltransferase, and the recently discovered spermine oxidase in the kinetic studies. In vitro studies confirmed that methylation did not protect spermine analogs from degradation, whereas the spermidine analog was stable. Both ␣-methylspermidine and bis-␣-methylspermine overcame the proliferative block of early liver regeneration in transgenic rats and reversed the cytostasis induced by an inhibition of ornithine decarboxylase in cultured fetal fibroblasts.Although the requirement of the natural polyamines spermidine, spermine, and their precursor putrescine for the growth of mammalian cells is extremely well documented, their specific functions in proliferative processes are largely unknown (1). Some of the published data appear to assign a central role to spermidine, whereas putrescine is supposed to serve as its precursor and spermine as a storage pool convertible back to spermidine. For the elucidation of the physiological roles of individual polyamines, metabolically stable derivatives of polyamines fulfilling their specific cellular functions would be extremely valuable. Methyl derivatives of spermidine and spermine have been used as substitutes for the natural polyamines both in vitro and in vivo. ␣-Methylspermidine (MeSpd), 1 ␣-methylspermine (MeSpm), and bis-␣-methylspermine (1,12-dimethylspermine, Me 2 Spm) are equally effective as the natural polyamines in inducing the conversion of right-handed B-DNA to left-handed Z-DNA (2). In addition to spermidine and spermine, cytostasis that resulted from the inhibition of the S-adenosylmethionine decarboxylase can be reversed by MeSpd but not by Me 2 Spm (3). Spermidine, spermine (because of its conversion to spermidine), and MeSpd serve as substrates for the synthesis of deoxyhypusine (an integral part of eukaryotic initiation factor 5A), whereas Me 2 Spm does not (3). Interestingly, all of the mentioned methylated derivatives of spermidine and spermine have been reported to reverse the cytostasis induced by difluoromethylornithine, a specific inhibitor of mammalian ornithine decarboxyl...

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Section: Discussionmentioning

confidence: 99%

“…Spermidine is not degraded at all, but monoethylspermine is as a good substrate for SMO as is spermine (21, 22). SMO, although inhibited to some extent, is much more resistant to the PAO inhibitor MDL72527 (21).…”

Section: Discussionmentioning

confidence: 99%

Metabolic Stability of α-Methylated Polyamine Derivatives and Their Use as Substitutes for the Natural Polyamines

Järvinen

Grigorenko

Khomutov

et al. 2005

Journal of Biological Chemistry

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“…due to acetylation) and/or oxidative events emanating from analog induction of polyamine-directed oxidases (Vujcic et al, 2002. Previously, we reported that induction of SSAT and related oxidative events are directly linked to cytochrome c release, caspase-3 activation and apoptosis induced by DENSPM .…”

Section: Discussionmentioning

confidence: 99%

Loss of inhibitor of apoptosis proteins as a determinant of polyamine analog-induced apoptosis in human melanoma cells

Chen

Kramer

et al. 2003

Oncogene

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We have previously shown that the clinically relevant polyamine analog N 1 ,N 11 -diethylnorspermine (DENSPM) causes rapid apoptosis in human melanoma SK-MEL-28 cells via a series of events that include mitochondrial release of cytochrome c and activation of the caspase cascade. Upstream to these events, DENSPM downregulates polyamine biosynthesis and potently upregulates polyamine catabolism at the level of spermidine/spermine N 1 -acetyltransferase (SSAT). In searching for downstream effectors that either contribute to or abrogate the apoptotic response, we observed that DENSPM treatment of SK-MEL-28 cells for 30 h led to cytosolic release of Smac/Diablo, a mitochondrial protein known to bind and inhibit the function of inhibitor of apoptosis proteins (IAPs). Subsequently, we found that DENSPM markedly lowered survivin and ML-IAP protein (but not XIAP) levels by 18 h via an apparently Smac/Diablo-independent pathway. Proteasome inhibitors fully prevented survivin and ML-IAP protein loss as well as apoptosis, suggesting that the proteasome-mediated degradation of survivin and ML-IAP is causally linked to the cellular outcome. We also observed that structural analogs of DENSPM which differentially induced SSAT and apoptosis lowered survivin and ML-IAP levels in a manner that correlated with enzyme activity. The linkage between IAPs and SSAT was more directly established by the finding that selective prevention of SSAT induction by small interfering RNA prevented survivin and ML-IAP loss as well as apoptosis during DENSPM treatment. Among the melanoma cell lines (SK-MEL-28, MALME-3M, A375 and LOX), survivin degradation correlated temporally with the onset of DENSPM induced apoptosis or growth inhibition. By contrast, ML-IAP degradation occurred only during rapid apoptosis seen in SK-MEL-28 cells. These data suggest a sequence of events whereby DENSPM induction of SSAT leads to loss of IAP proteins and a more fulminate apoptotic response. The findings implicate survivin and ML-IAP as important determinants of polyamine analog drug action in melanoma cells.

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“…Polyamine oxidation can occur by acetylation of spermine or spermidine by spermidine/spermine N 1 -acetyltransferase prior to back-conversion by acetyl polyamine oxidase (17,18) or by direct conversion of spermine to spermidine by polyamine oxidase 1 (PAO1), also called spermine oxidase (19,20). We have recently demonstrated that H. pylori up-regulates expression and activity of PAO1 in macrophages, which acts to regulate intracellular spermine levels in activated cells (13).…”

Section: Specific Knockdown Of Odc By Sirna Increases H Pyloristimulmentioning

confidence: 99%

Spermine Causes Loss of Innate Immune Response to Helicobacter pylori by Inhibition of Inducible Nitric-oxide Synthase Translation

Bussière¹,

Chaturvedi

Cheng

et al. 2005

Journal of Biological Chemistry

126

147

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Helicobacter pylori infection of the stomach elicits a vigorous but ineffective host immune and inflammatory response, resulting in persistence of the bacterium for the life of the host. We have reported that in macrophages, H. pylori up-regulates inducible NO synthase (iNOS) and antimicrobial NO production, but in parallel there is induction of arginase II, generating ornithine, and of ornithine decarboxylase (ODC), generating polyamines. Spermine, in particular, has been shown to restrain immune response in activated macrophages by inhibiting proinflammatory gene expression. We hypothesized that spermine could prevent the antimicrobial effects of NO by inhibiting iNOS in macrophages activated by H. pylori. Spermine did not affect the upregulation of iNOS mRNA levels but in a concentrationdependent manner significantly attenuated iNOS protein levels and NO production. Reduction in iNOS protein was due to inhibition of iNOS translation and not due to iNOS degradation. ODC knockdown with small interfering (si) RNA resulted in increased H. pylori-stimulated iNOS protein expression and NO production without altering iNOS mRNA levels. When macrophages were cocultured with H. pylori, killing of bacteria was enhanced by transfection of ODC siRNA and prevented by addition of spermine. These results identify a mechanism of immune dysregulation induced by H. pylori in which stimulated spermine synthesis by the arginase-ODC pathway inhibits iNOS translation and NO production, leading to persistence of the bacterium and risk for peptic ulcer disease and gastric cancer.Helicobacter pylori is a Gram-negative, microaerophilic bacterium that selectively colonizes the human stomach. Current prevalence of H. pylori is ϳ30 -40% of the population in the United States (1) and substantially higher in underdeveloped regions. H. pylori infection induces a vigorous mucosal immune response that fails to eradicate the organism and results in chronic gastritis that can lead to peptic ulcers and gastric cancer. In addition to a chronic lymphocytic response, H. pylori infection induces activation of an innate immune response in neutrophils, monocytes, and macrophages (2-8). Inducible NO synthase (iNOS) 1 -derived NO is a central effector molecule in the innate immune response to pathogens, with essential antimicrobial functions in host defense. We have reported that H. pylori induces iNOS expression and activity in macrophages (4 -7). H. pylori is considered a noninvasive pathogen, but it can disrupt epithelial integrity, and its antigens are present in the lamina propria (3). H. pylori can induce iNOS and other innate immune response genes in macrophages even when separated by filter supports or when water extracts are used (6). Although H. pylori-induced NO production can kill the bacterium in vitro (7, 9), it survives in the stomach, despite detection of iNOS in infected gastric mucosa (10).Production of NO by macrophages can be limited by H. pylori arginase that competes with iNOS for the same substrate, L-arginine (7) under conditions ...

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Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin

Cited by 197 publications

References 39 publications

Metabolic Stability of α-Methylated Polyamine Derivatives and Their Use as Substitutes for the Natural Polyamines

Metabolic Stability of α-Methylated Polyamine Derivatives and Their Use as Substitutes for the Natural Polyamines

Loss of inhibitor of apoptosis proteins as a determinant of polyamine analog-induced apoptosis in human melanoma cells

Spermine Causes Loss of Innate Immune Response to Helicobacter pylori by Inhibition of Inducible Nitric-oxide Synthase Translation

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