Effects of inhibitors of spermidine synthase and spermine synthase on polyamine synthesis in rat tissues

Shirahata, Akira; Takahashi, Norio; Beppu, Takanobu; Hosoda, Harumi; Samejima, Keijiro

doi:10.1016/0006-2952(93)90449-7

Cited by 56 publications

(49 citation statements)

References 22 publications

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“…The IC 50 value was 0.2 µM, which compares well with values of 0.1-0.4 µM under similar assay conditions for other PAPTs 2,15 . Spermidine synthesis by TmPAPT had an optimal pH of 7.5 and was greatly increased at higher temperatures, with the k cat value increasing from 0.08 s −1 at 37 °C to 3.3 s −1 at 90 °C.…”

Section: Papt Enzymatic Activity and Polyamine Synthesissupporting

confidence: 79%

The crystal structure of spermidine synthase with a multisubstrate adduct inhibitor

Korolev

Ikeguchi

Skarina³

et al. 2001

Nat. Struct Biol.

134

164

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Polyamines are essential in all branches of life. Spermidine synthase (putrescine aminopropyltransferase, PAPT) catalyzes the biosynthesis of spermidine, a ubiquitous polyamine. The crystal structure of the PAPT from Thermotoga maritima (TmPAPT) has been solved to 1.5 Å resolution in the presence and absence of AdoDATO (S-adenosyl-1,8-diamino-3-thiooctane), a compound containing both substrate and product moieties. This, the first structure of an aminopropyltransferase, reveals deep cavities for binding substrate and cofactor, and a loop that envelops the active site. The AdoDATO binding site is lined with residues conserved in PAPT enzymes from bacteria to humans, suggesting a universal catalytic mechanism. Other conserved residues act sterically to provide a structural basis for polyamine specificity. The enzyme is tetrameric; each monomer consists of a C-terminal domain with a Rossmann-like fold and an Nterminal β-stranded domain. The tetramer is assembled using a novel barrel-type oligomerization motif.The nearly ubiquitous polyamines (putrescine, spermidine and spermine) are polycationic mediators of cell proliferation and differentiation 1 whose functions likely provide both stability and neutralization for nucleic acids. The abundance of polyamines is tightly regulated through biosynthesis, degradation, uptake and efflux 2 . The biosynthesis of polyamines is carried out by three highly conserved polyamine biosynthetic enzymes ( Fig. 1): ornithine decarboxylase, putrescine amino-propyltransferase (PAPT) and spermidine aminopropyltransferase (SAPT). The strong correlation of polyamine synthesis with cell growth renders these aminopropyl transferases (APTs) attractive targets for the development of antiproliferative therapeutics. The APTs are known to be inhibited by their common nucleoside product, as well as other nucleoside analogs. Most inhibitors, such as the nucleoside-polyamine adduct 4 , is also a potent inhibitor of PAPT.The catalytic mechanism of polyamine biosynthesis is unclear, partly because there is no detailed structural information for this class of enzymes. To provide structural insights into the mechanism of spermidine synthesis, the crystal structure of PAPT from Thermotoga maritima (TmPAPT) was determined at high resolution in ligand-free and AdoDATO-bound states. These structures provide insight into the function of the enzyme and its potential mechanism of action. The identification of conserved active site residues should enable accurate models of the mammalian enzymes. The PAPT foldThe structure of TmPAPT was solved using multiwavelength anomalous diffraction (MAD) from selenomethionine (SeMet)-containing crystals. The TmPAPT monomer consists of two domains: an N-terminal domain, composed of six β-strands, and a Rossmann-like C-terminal domain (Fig. 2). The larger C-terminal catalytic core domain (residues 75-296) consists of a seven-stranded β-sheet with a strand order of 9↑, 8↑, 7↑, 10↑, 11↑, 13↓, 12↑ flanked by nine α-helices. This domain resembles a topology observed in...

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Section: Papt Enzymatic Activity and Polyamine Synthesissupporting

confidence: 79%

The crystal structure of spermidine synthase with a multisubstrate adduct inhibitor

Korolev

Ikeguchi

Skarina³

et al. 2001

Nat. Struct Biol.

134

164

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“…In cultured cells, inhibition of spd-syn led to a decrease of spd and an increase of spm, but in short-term experiments, using the inhibitors, S-adenosyl-1,8-diamino-3-thiooctane (AdoDato) 7) and trans-4-methylcyclohexylamine (4MCHA), 8) little or no change in growth was observed. We obtained similar results in a study using rats administered 4MCHA, dissolved in drinking water and available ad libitum, for a period of 10 d. 9) The basic response of cells treated with spdsyn inhibitors was elucidated from these studies. However, no detailed quantitative study has been performed in vivo on the effect of spd-syn inhibitors, although some studies have used cyclohexylamine, 10) which is an order of magnitude weaker than 4MCHA.…”

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confidence: 59%

“…11) The following stable isotopelabeled polyamines were prepared in our laboratory 12) : [1,[4][5][6][7][8][9][10][11][12][13][14][15] N-spm). Standard polyamines were purchased from Sigma (Japan).…”

Section: Methodsmentioning

confidence: 99%

Control of Spermidine and Spermine Levels in Rat Tissues by trans-4-Methylcyclohexylamine, a Spermidine-Synthase Inhibitor

Kobayashi

Watanabe

et al. 2005

Biological & Pharmaceutical Bulletin

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The polyamines, putrescine, spermidine (spd) and spermine (spm), present in mammalian tissues, are regulated under a variety of physiological conditions. Their tissue concentrations are controlled by a number of factors: (1) polyamine biosynthetic enzymes, such as ornithine decarboxylase (ODC), S-adenosyl-L-methionine decarboxylase (AdoMetDC), spermidine synthase (spd-syn) and spermine synthase (spm-syn); (2) polyamine catabolic enzymes, such as spd/spm N 1 -acetyltransferase (SSAT), polyamine oxidase (PAO), spermine oxidase (SMO) and diamine oxidase

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“…These use radioisotope (RI), 10 high-performance liquid chromatography (HPLC), 11,12 homogeneous time-resolved fluorescence 13 or mass spectrometry 14 methods. The RI method with 35 S-dcSAM as a substrate for Apt activity measurement needs preparation of 35 S-dcSAM by enzymatic reaction with S-adenosylmethionine (SAM) synthase, ATP, radio isotope-labeled methionine and SAM decarboxylase, followed by manual column chromatographic purification because dcSAM is not commercially available.…”

Section: Introductionmentioning

confidence: 99%

Determination of Cellular Aminopropyltransferase Activity Using Precolumn Fluorescent Etheno-Derivatization with High-Performance Liquid Chromatography

et al. 2012

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Effects of inhibitors of spermidine synthase and spermine synthase on polyamine synthesis in rat tissues

Cited by 56 publications

References 22 publications

The crystal structure of spermidine synthase with a multisubstrate adduct inhibitor

The crystal structure of spermidine synthase with a multisubstrate adduct inhibitor

Control of Spermidine and Spermine Levels in Rat Tissues by trans-4-Methylcyclohexylamine, a Spermidine-Synthase Inhibitor

Determination of Cellular Aminopropyltransferase Activity Using Precolumn Fluorescent Etheno-Derivatization with High-Performance Liquid Chromatography

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