Crystal Structure of Plasmodium falciparum Spermidine Synthase in Complex with the Substrate Decarboxylated S-adenosylmethionine and the Potent Inhibitors 4MCHA and AdoDATO

Dufe, Veronica Tamu; Qiu, Wei; Müller, Ingrid; Hui, Raymond; Walter, Rolf D.; Al-Karadaghi, Salam

doi:10.1016/j.jmb.2007.07.053

Cited by 53 publications

(79 citation statements)

References 38 publications

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“…The SPDS gene is widely conserved in a variety of organisms and encodes a compact globular protein (Dufe et al, 2007). A number of diverse plants possess unique motifs that are attached to the 59 end of the open reading frame that encodes the conserved enzymatic portion of the protein (Haider et al, 2005;Dufe et al, 2007).…”

Section: Development Is Arrested After Spds Silencingmentioning

confidence: 99%

Spermidine Is a Morphogenetic Determinant for Cell Fate Specification in the Male Gametophyte of the Water FernMarsilea vestita

2010

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Here, we show that the polyamine spermidine plays a key role as a morphogenetic determinant during spermatid development in the water fern Marsilea vestita. Spermidine levels rise first in sterile jacket cells and then increase dramatically in spermatogenous cells as the spermatids mature. RNA interference and drug treatments were employed to deplete spermidine in the gametophyte at different stages of gametogenesis. Development in spermidine-depleted gametophytes was arrested before the completion of the last round of cell divisions. In spermidine-depleted spermatogenous cells, chromatin failed to condense properly, basal body positioning was altered, and the microtubule ribbon was in disarray. When cyclohexylamine, a spermidine synthase (SPDS) inhibitor, was added at the start of spermatid differentiation, the spermatid nuclei remained round, centrin failed to localize into basal bodies, thus blocking basal body formation, and the microtubule ribbon was completely abolished. In untreated gametophytes, spermidine made in the jacket cells moves into the spermatids, where it is involved in the unmasking of stored SPDS mRNAs, leading to substantial spermidine synthesis in the spermatids. We found that treating spores directly with spermidine or other polyamines was sufficient to unmask a variety of stored mRNAs in gametophytes and arrest development. Differences in patterns of transcript distribution after these treatments suggest that specific transcripts reside in different locations in the dry spore; these differences may be linked to the timing of unmasking and translation for that mRNA during development.

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Section: Development Is Arrested After Spds Silencingmentioning

confidence: 99%

Spermidine Is a Morphogenetic Determinant for Cell Fate Specification in the Male Gametophyte of the Water FernMarsilea vestita

2010

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“…Some aminopropyltransferases such as human spermidine synthase (SpdS) (which acts upon putrescine) and spermine synthase (SpmS) (acting on spermidine) are highly specific for their amine acceptors, [12][13][14] while others, such as those from acute thermophiles, which contain a variety of polyamines not found in mammals, are less discriminating. 12,13,[15][16][17] There are now numerous published structures for aminopropyltransferases including those for SpdS from Thermotoga maritima (TmSpdS), 16 Caenorhabditis elegans, 18 Plasmodium falciparum (PfSpdS), 19 Helicobacter pylori, 20 human (hSpdS), 13 Arabidopsis thaliana (PDB code 2Q41), and Trypanosoma cruzi (PDB code 3BWC), aminopropylagmatine/aminopropylcadaverine synthases from Thermus thermophilus (PDB code 1UIR), Pyrococcus horikoshii (PDB code 2ZSU) and Pyrococcus furiosus, 15 and SpmS from humans (hSpmS). 14 A general mechanism for aminopropyl transfer has been proposed based on kinetic studies of hSpdS, TmSpdS, and hSpmS, their structures with bound substrates and inhibitors, and the results of site-directed mutagenesis of key residues (Fig.…”

Section: Introductionmentioning

confidence: 99%

Binding and inhibition of human spermidine synthase by decarboxylated S‐adenosylhomocysteine

Šečkutė¹,

McCloskey²,

Thomas³

et al. 2011

Protein Science

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Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5 0 -deoxy-5 0 -methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dosedependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 Å resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K d of 1.1 6 0.3 lM in the absence of putrescine and 3.2 6 0.1 lM in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold.

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“…20 and 21), aminopropylagmatine synthase uses agmatine (decarboxylated arginine) (22), and thermospermine synthase adds the aminopropyl group to N-1 of SPD 5 (18). Structures have been determined by x-ray crystallography for a number of SpdSyns, including those from Thermotoga maritima (23), Caenorhabditis elegans (24), Plasmodium falciparum (25), Helicobacter pylori (26), and human (27), and for the aminopropylagmatine synthases from Thermus thermophilus (Protein Data Bank code 1UIR) (22) and Pyrococcus furiosus (28). A general mechanism for aminopropyl transfer has been proposed based on the human and T. maritima SpdSyn structures bound to substrates and inhibitors and the results of site-directed mutagenesis of key residues (27).…”

mentioning

confidence: 99%

Crystal Structure of Human Spermine Synthase

Min

Zeng

et al. 2008

Journal of Biological Chemistry

105

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The crystal structures of two ternary complexes of human spermine synthase (EC 2.5.1.22), one with 5-methylthioadenosine and spermidine and the other with 5-methylthioadenosine and spermine, have been solved. They show that the enzyme is a dimer of two identical subunits. Each monomer has three domains: a C-terminal domain, which contains the active site and is similar in structure to spermidine synthase; a central domain made up of four ␤-strands; and an N-terminal domain with remarkable structural similarity to S-adenosylmethionine decarboxylase, the enzyme that forms the aminopropyl donor substrate. Dimerization occurs mainly through interactions between the N-terminal domains. Deletion of the N-terminal domain led to a complete loss of spermine synthase activity, suggesting that dimerization may be required for activity. The structures provide an outline of the active site and a plausible model for catalysis. The active site is similar to those of spermidine synthases but has a larger substrate-binding pocket able to accommodate longer substrates. Two residues (Asp 201 and Asp 276 ) that are conserved in aminopropyltransferases appear to play a key part in the catalytic mechanism, and this role was supported by the results of site-directed mutagenesis. The spermine synthase⅐5-methylthioadenosine structure provides a plausible explanation for the potent inhibition of the reaction by this product and the stronger inhibition of spermine synthase compared with spermidine synthase. An analysis to trace possible evolutionary origins of spermine synthase is also described.Polyamines are essential for normal growth and development, and the polyamine biosynthetic pathway is an important target for the design of therapeutic agents (1, 2). The predominant polyamines in mammalian cells are spermidine (SPD) 4 and spermine (SPM). These polyamines are made by the sequential addition of aminopropyl groups from decarboxylated S-adenosylmethionine (dcAdoMet). The addition of the first aminopropyl group to the diamine precursor putrescine is catalyzed by spermidine synthase (SpdSyn). The addition of a second aminopropyl group to the N-10 position 5 of SPD to form SPM is catalyzed by spermine synthase (SpmSyn) (Fig. 1). The critical importance of polyamines is demonstrated by studies using mice with gene deletions of ornithine decarboxylase (3) or S-adenosylmethionine decarboxylase (AdoMetDC) (4), both of which are lethal at the earliest stages of embryonic development. No SpdSyn knock-out mice have been described, but studies in yeast show that this gene is required for viability and that SPD plays an essential role as a precursor of hypusine formed in the post-translational modification of eukaryotic initiation factor 5A (5). SpmSyn has received relatively little attention compared with the large number of publications describing ornithine decarboxylase, AdoMetDC, and SpdSyn (1, 6 -8). The SpmSyn gene is not essential for growth of yeast (9). Mutants lacking the AdoMetDC gene, which cannot make SPD or SPM, grow normally when ...

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Crystal Structure of Plasmodium falciparum Spermidine Synthase in Complex with the Substrate Decarboxylated S-adenosylmethionine and the Potent Inhibitors 4MCHA and AdoDATO

Cited by 53 publications

References 38 publications

Spermidine Is a Morphogenetic Determinant for Cell Fate Specification in the Male Gametophyte of the Water FernMarsilea vestita

Spermidine Is a Morphogenetic Determinant for Cell Fate Specification in the Male Gametophyte of the Water FernMarsilea vestita

Binding and inhibition of human spermidine synthase by decarboxylated S‐adenosylhomocysteine

Crystal Structure of Human Spermine Synthase

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