Guide Molecule-driven Stereospecific Degradation of α-Methylpolyamines by Polyamine Oxidase

Järvinen, Aki; Keinänen, Tuomo A.; Grigorenko, Nikolay A.; Khomutov, Alex R.; Uimari, Anne; Vepsäläinen, Jouko; Närvänen, Ale; Alhonen, Leena; Jänne, Juhani

doi:10.1074/jbc.m509959200

Cited by 21 publications

(29 citation statements)

References 38 publications

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“…We recently reported that mammalian polyamine oxidase, normally using achiral substrates, displays cryptic stereospecificity when ␣-methylated polyamines are used as the substrates (32,33). In fact, the latter results (33) indicate that polyamine oxidase has a k cat /K m value (efficiency of the enzyme catalysis) for the (S,S)-isomer of Me 2 Spm that is nearly 30 times higher than that for the (R,R)-isomer. However, this difference may not be practically relevant as non-acetylated polyamines are poor substrates for mammalian polyamine oxidase in the absence of aldehydes.…”

Section: Tablementioning

confidence: 90%

Role of Hypusinated Eukaryotic Translation Initiation Factor 5A in Polyamine Depletion-induced Cytostasis

Hyvönen

Keinänen

Cerrada-Gimenez

et al. 2007

Journal of Biological Chemistry

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We have earlier shown that ␣-methylated spermidine and spermine analogues rescue cells from polyamine depletion-induced growth inhibition and maintain pancreatic integrity under severe polyamine deprivation. However, because ␣-methylspermidine can serve as a precursor of hypusine, an integral part of functional eukaryotic translation initiation factor 5A required for cell proliferation, and because ␣,-bismethylspermine can be converted to methylspermidine, it is not entirely clear whether the restoration of cell growth is actually attributable to hypusine formed from these polyamine analogues. Here, we have used optically active isomers of methylated spermidine and spermine and show that polyamine depletion-induced acute cytostasis in cultured cells could be reversed by all the isomers of the methylpolyamines irrespective of whether they served or not as precursors of hypusine. In transgenic rats with activated polyamine catabolism, all the isomers similarly restored liver regeneration and reduced plasma ␣-amylase activity associated with induced pancreatitis. Under the above experimental conditions, the (S,S)-but not the (R,R)-isomer of bismethylspermine was converted to methylspermidine apparently through the action of spermine oxidase strongly preferring the (S,S)-isomer. Of the analogues, however, only (S)-methylspermidine sustained cell growth during prolonged (more than 1 week) inhibition of polyamine biosynthesis. It was also the only isomer efficiently converted to hypusine, indicating that deoxyhypusine synthase likewise possesses hidden stereospecificity. Taken together, the results show that growth inhibition in response to polyamine depletion involves two phases, an acute and a late hypusine-dependent phase.A large number of studies have indicated that a continuous supply of the polyamines (spermidine and spermine) is required for animal cell proliferation to occur as polyamine depletion resulted either from a specific inhibition of their biosynthesis (1) or from an activation of their catabolism (2) invariably leads to growth inhibition. The molecular mechanisms involved in the requirement of polyamines for animal cell growth are largely unknown besides the fact that spermidine, but not spermine, serves as the sole biosynthetic precursor for hypusine, an unusual amino acid that is an integral component of eukaryotic translation initiation factor 5A (eIF5A) 4 (3). Because functional (hypusinated) eIF5A is required for animal cell growth (4, 5), it is often difficult to judge whether spermidine depletion-induced growth inhibition is secondary to hypusine deprivation. The finding that cytostasis resulting from an inhibition of S-adenosylmethionine decarboxylase in cultured cells is reversed by ␣-methylspermidine (MeSpd) but not by ␣,-bismethylspermine (Me 2 Spm) indicates that growth inhibition was attributable to hypusine depletion as MeSpd, but not Me 2 Spm, can serve as the biosynthetic precursor for hypusine formation (6). On the other hand, MeSpd and both singly and doubly methylated spermine deri...

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Section: Tablementioning

confidence: 90%

Role of Hypusinated Eukaryotic Translation Initiation Factor 5A in Polyamine Depletion-induced Cytostasis

Hyvönen

Keinänen

Cerrada-Gimenez

et al. 2007

Journal of Biological Chemistry

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“…Similarly, SPD was a major metabolite with APAO, but some other competing pathways were also detected. Interestingly, with charge retaining EtSPM and DESPM the reaction velocity is slower when compared with N 1 -AcSPM and N 1 ,N 12 -DiAcSPM, respectively (Järvinen et al 2006a). BnSPM was metabolized to SPD by both oxidases, APAO being more efficient than SMO.…”

Section: Discussionmentioning

confidence: 99%

“…SMO (PAOh1) was initially cloned and characterized in 2001 (Wang et al 2001). Cloning of both the enzymes has made them readily available for recombinant protein production that has facilitated further testing of their substrate properties (Järvinen et al 2006a; Wang et al 2003; Wang et al 2005a). During polyamine catabolism, SPM and SPD are first acetylated by spermidine/spermine N 1 -acetyltransferase (SSAT) and subsequently oxidized by APAO to produce SPD and PUT, respectively, and to yield stoichiometric amounts of hydrogen peroxide and 3-acetamidopropanal.…”

Section: Introductionmentioning

confidence: 99%

“…APAO prefers acetylated polyamines over non-acetylated polyamines, and its natural substrates are N 1 ,N 12 - DiAcSPM, N 1 - AcSPM and N 1 - AcSPD (Seiler 1995; Wang et al 2005a). In addition to the natural polyamines, some terminally N -alkylated polyamines are degraded by APAO, and some of the otherwise metabolically stable polyamine analogues are catabolized when certain aldehydes are present in the reaction mixture (Järvinen et al 2005; Järvinen et al 2006a; Vujcic et al 2003; Wu et al 2003). By contrast, SMO possesses very different substrate specificities for the natural polyamines as compared with APAO.…”

Section: Introductionmentioning

confidence: 99%

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Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

et al. 2009

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SUMMARYN-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N, N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N'-(3-ethylaminopropyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD (K m(APAO) =10μM, k cat(APAO) =1.1s −1 and K m(SMO) =28μM, k cat(SMO) =0.8s −1 , respectively), metabolized BnEtSPM to EtSPD (K m(APAO) =0.9 μM, k cat(APAO) =1.1s −1 and K m(SMO) =51μM, k cat(SMO) =0.4s −1 , respectively), and metabolized DBSPM to BnSPD (K m(APAO) =5.4μM, k cat(APAO) = 2.0s −1 and K m(SMO) =33μM, k cat(SMO) =0.3s −1 , respectively). Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD (EtSPM K m(APAO) =16μM, k cat(APAO) =1.5s −1 ; K m(SMO) =25μM, k cat(SMO) =8.2s −1 ; BnSPM K m(APAO) =6.0μM, k cat(APAO) =2.8s −1 ; K m(SMO) =19μM, k cat(SMO) =0.8s −1 , respectively). Surprisingly, E t S P D ( K m(APAO) =37μM, k cat(APAO) =0.1s −1 ; K m(SMO) =48μM, k cat(SMO) =0.05s −1 ) and BnSPD (K m(APAO) =2.5μM, k cat(APAO) =3.5s −1 ; K m(SMO) =60μM, k cat(SMO) =0.54s −1 ) were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 hours of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could play an important role in polyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

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“…More recent studies, in which (S)- and (R)- isomers of α-methylspermidine and (S,S)-, (R,S)-, and (R,R)-diastereomers of α,α′-(dimethyl)spermine have been used, have confirmed and extended these important findings (67, 191–196). Such studies have shown the maintenance of nucleic acid structure (196, 197), the stereospecificity of oxidation reactions (192, 194), the ability of derivatives that can support hypusine synthesis to support growth (67, 188, 190), and the role of polyamines in other growth processes including hair synthesis (198), liver regeneration, and adipocyte differentiation (199). Of particular interest is the ability to overcome acute liver damage and pancreatitis related to polyamine depletion by provision of α-methylspermidine or α,α′-(dimethyl)spermine (168, 193, 195, 200, 201).…”

Section: Polyamine Analogs and Derivatives As Research Tools And mentioning

confidence: 99%

Current Status of the Polyamine Research Field

Pegg

Casero

2011

Methods in Molecular Biology

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175

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This chapter provides an overview of the polyamine field and introduces the 32 other chapters that make up this volume. These chapters provide a wide range of methods, advice, and background relevant to studies of the function of polyamines, the regulation of their content, their role in disease, and the therapeutic potential of drugs targeting polyamine content and function. The methodology provided in this new volume will enable laboratories already working in this area to expand their experimental techniques and facilitate the entry of additional workers into this rapidly expanding field.

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Guide Molecule-driven Stereospecific Degradation of α-Methylpolyamines by Polyamine Oxidase

Cited by 21 publications

References 38 publications

Role of Hypusinated Eukaryotic Translation Initiation Factor 5A in Polyamine Depletion-induced Cytostasis

Role of Hypusinated Eukaryotic Translation Initiation Factor 5A in Polyamine Depletion-induced Cytostasis

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

Current Status of the Polyamine Research Field

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