Polyamine synthesis in mammalian tissues. Isolation and characterization of spermidine synthase from bovine brain

Raina, Aarne; Hyvönen, Tapani; Eloranta, Terho O.; Voutilainen, M; Samejima, Keijiro; Yamanoha, Banri

doi:10.1042/bj2190991

Cited by 40 publications

(23 citation statements)

References 38 publications

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“…The decrease in spermidine concentration found in the meu1⌬ cells could have been due to inhibition of spermidine synthase by methylthioadenosine as has been described for spermidine synthase from other sources (30)(31)(32). By using crude extracts from MEU1 ϩ and meu1⌬ cultures, we found that this was indeed the case (Fig.…”

Section: Odc Is More Stable In Meu1⌬supporting

confidence: 56%

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Studies on the regulation of ornithine decarboxylase in yeast: Effect of deletion in the MEU1 gene

Chattopadhyay

Tabor

2005

Proc. Natl. Acad. Sci. U.S.A.

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Methylthioadenosine is formed during the biosynthesis of spermidine and of spermine and is metabolized by methylthioadenosine phosphorylase, an enzyme missing in several tumor cell lines. In Saccharomyces cerevisiae, this enzyme is coded by the MEU1 gene. We have now studied the effect of the meu1 deletion on polyamine metabolism in yeast. We found that the effects of the meu1⌬ mutation mostly depend on the stage of cell growth. As the cell density increases, there is a marked fall in the level of ornithine decarboxylase (ODC) in the MEU1 ؉ cells, which we show is caused by an antizyme-requiring degradation system. In contrast, there is only a small decrease in the ODC level in the meu1⌬ cells. The meu1⌬ cells have a higher putrescine and a lower spermidine level than MEU1 ؉ cells, suggesting that the decreased spermidine level in the meu1⌬ cultures is responsible for the greater apparent stability of ODC in the meu1⌬ cells. The lower spermidine level in the meu1⌬ cells probably results from an inhibition of spermidine synthase by the methylthioadenosine that presumably accumulates in these mutants. In both MEU1 ؉ and the meu1⌬ cultures, the ODC levels were markedly decreased by the addition of spermidine to the media, and thus our results contradict the postulation of Subhi et al. methylthioadenosine ͉ methylthioadenosine phosphorylase ͉ polyamines ͉ spermidine ͉ putrescine O rnithine decarboxylase (ODC) is the key regulatory enzyme of the polyamine biosynthetic pathway and catalyzes the decarboxylation of ornithine to 1,4-diaminobutane (putrescine). Putrescine is then converted to spermidine and spermine by the addition of aminopropyl groups from decarboxylated Sadenosylmethionine (1). Methylthioadenosine is formed as a byproduct of these aminopropyl transferase reactions (spermidine synthase and spermine synthase) (2-4) and is then converted to adenine and to methionine by the ''methionine salvage'' pathway (5-9). The first step in the methionine salvage pathway is the phosphorolysis of methylthioadenosine to methylthioribose-1-phosphate and adenine by methylthioadenosine phosphorylase (EC 2.4.2.28). The metabolism of methylthioadenosine in mammalian cells has been of special interest because methylthioadenosine phosphorylase is absent in many tumors (10)(11)(12)(13)(14)(15). Thomas et al. (16) and Subhi et al. (17) have shown that, in Saccharomyces cerevisiae, methylthioadenosine phosphorylase is coded by the MEU1 gene and have described meu1⌬ mutants. The availability of these yeast mutants deleted in the methylthioadenosine phosphorylase gene permits more extensive studies on the role of this gene and the methionine salvage pathway in polyamine metabolism in yeast, and a comparison of these effects in yeast with those reported in mammalian cells. Therefore, as an extension of our work on the regulation of ornithine decarboxylase and of polyamine pools in yeast (18), we have now studied the effect of the meu1⌬ deletion on these parameters. Our findings were different from those reported by Subhi et al.

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Section: Odc Is More Stable In Meu1⌬supporting

confidence: 56%

“…6). Spermidine synthases from rat prostate and bovine brain were also inhibited in an in vitro system by methylthioadenosine (30)(31)(32). However, we have not been able to show any change in the ODC activity or the putrescine or spermidine content when either MEU1 ϩ or meu1⌬ cells were grown with added methylthioadenosine.…”

Section: Discussionmentioning

confidence: 64%

Studies on the regulation of ornithine decarboxylase in yeast: Effect of deletion in the MEU1 gene

Chattopadhyay

Tabor

2005

Proc. Natl. Acad. Sci. U.S.A.

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“…We assumed in the model that 5ЈMTA was virtually zero. This assumption was supported by the fact that 5ЈMTA is quickly removed by 5ЈMTA phosphorylase (47,48). Table 2 gives the differential equations for the different metabolites and other time-dependent variables.…”

Section: Study System Definition and Mathematical Model Description-mentioning

confidence: 80%

Mathematical Modeling of Polyamine Metabolism in Mammals

Rodríguez‐Caso

Montañez

Cascante

et al. 2006

Journal of Biological Chemistry

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Polyamines are considered as essential compounds in living cells, since they are involved in cell proliferation, transcription, and translation processes. Furthermore, polyamine homeostasis is necessary to cell survival, and its deregulation is involved in relevant processes, such as cancer and neurodegenerative disorders. Great efforts have been made to elucidate the nature of polyamine homeostasis, giving rise to relevant information concerning the behavior of the different components of polyamine metabolism, and a great amount of information has been generated. However, a complex regulation at transcriptional, translational, and metabolic levels as well as the strong relationship between polyamines and essential cell processes make it difficult to discriminate the role of polyamine regulation itself from the whole cell response when an experimental approach is given in vivo. To overcome this limitation, a bottom-up approach to model mathematically metabolic pathways could allow us to elucidate the systemic behavior from individual kinetic and molecular properties. In this paper, we propose a mathematical model of polyamine metabolism from kinetic constants and both metabolite and enzyme levels extracted from bibliographic sources. This model captures the tendencies observed in transgenic mice for the so-called key enzymes of polyamine metabolism, ornithine decarboxylase, S-adenosylmethionine decarboxylase and spermine spermidine N-acetyl transferase. Furthermore, the model shows a relevant role of S-adenosylmethionine and acetyl-CoA availability in polyamine homeostasis, which are not usually considered in systemic experimental studies.During much of the last century, biology was an attempt to reduce biological phenomena to the behavior of molecules. Despite the enormous success of this approach, most biological functions arise from interactions among many components, yielding nonlinear behavior that has been fine tuned by natural selection to achieve specific functional properties (1, 2). Therefore, a comprehensive understanding of biological functions requires a new systemic perspective. In the last few years, systems biology and synthetic biology have emerged to fulfill this goal (3-5). Systems biology approaches are hypothesis-driven and involve iterative rounds of model building, prediction, experimentation, and model refinement and development (6, 7). Computer science development is allowing faster and faster numerical simulations of mathematical models. Interesting and relevant mathematical models of several well known metabolic pathways have been published very recently (8 -10). Far from replacing knowledge acquisition from experimental evidence, mathematical modeling allows to test and define more accurately hypothesis that have to be experimentally tested later. Furthermore, modeling allows to build a comprehensive framework based on a compilation of the experimental data provided by the scientific community. With this philosophy, we propose and study a mathematical model of polyamine metabolism. We...

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“…Many small molecule (65)(66)(67)(68) and DNA methyltransferases (69,70) are monomers, especially those isolated from nonmammalian sources. Spermine and spermidine synthase, on the other hand, appear to be dimers of identical subunits (59,61,62). The data are less conclusive in the case of nicotianamine synthase.…”

Section: Resultsmentioning

confidence: 54%

“…Spermidine synthase from bovine brain was purified 570-fold utilizing S-adenosyl-(5Ј)-3-thiopropylamine (decarboxylated AdoHcy)-Sepharose as the adsorbent. In this gel, the ligand forms a sulfonium linkage to the solid support by displacing the bromide of bromoacetyl-Sepharose (59). A similar coupling strategy was employed in the synthesis of AdoHcy-Sepharose (37).…”

Section: Resultsmentioning

confidence: 99%

Purification, Characterization, and Cloning of an S-Adenosylmethionine-dependent 3-Amino-3-carboxypropyltransferase in Nocardicin Biosynthesis

Reeve

Breazeale

Townsend

1998

Journal of Biological Chemistry

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S-Adenosylmethionine:nocardicin 3-amino-3-carboxypropyltransferase catalyzes the biosynthetically rare transfer of the 3-amino-3-carboxypropyl moiety from S-adenosylmethionine to a phenolic site in the ␤-lactam substrates nocardicin E, F, and G, a late step of the biosynthesis of the monocyclic ␤-lactam antibiotic nocardicin A. Whereas a number of conventional methods were ineffective in purifying the transferase, it was successfully obtained by two complementary affinity chromatography steps that took advantage of the two substrate-two product reaction scheme. S-Adenosylhomocysteine-agarose selected enzymes that utilize S-adenosylmethionine, and a second column, nocardicin Aagarose, specifically bound the desired transferase to yield the enzyme as a single band of 38 kDa on a silverstained SDS-polyacrylamide gel. The transferase is active as a monomer and exhibits sequential kinetics. Further kinetic characterization of this protein is described and its role in the biosynthesis of nocardicin A discussed. The gene encoding this transferase was cloned from a sublibrary of Nocardia uniformis DNA. Translation gave a protein of deduced mass 32,386 Da which showed weak homology to small molecule methyltransferases. However, three correctly disposed signature motifs characteristic of these enzymes were observed.

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Polyamine synthesis in mammalian tissues. Isolation and characterization of spermidine synthase from bovine brain

Cited by 40 publications

References 38 publications

Studies on the regulation of ornithine decarboxylase in yeast: Effect of deletion in the MEU1 gene

Studies on the regulation of ornithine decarboxylase in yeast: Effect of deletion in the MEU1 gene

Mathematical Modeling of Polyamine Metabolism in Mammals

Purification, Characterization, and Cloning of an S-Adenosylmethionine-dependent 3-Amino-3-carboxypropyltransferase in Nocardicin Biosynthesis

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