Specificity of mammalian spermidine synthase and spermine synthase

Pegg, Anthony E.; Shuttleworth, Kay L.; Hibasami, Hiroshige

doi:10.1042/bj1970315

Cited by 59 publications

(24 citation statements)

References 33 publications

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“…The mammalian systems that have been investigated, however, show that active OmDCase decreases in response to additions of putrescine (27) and the putrescine analog, 1,3-diaminopropane (28,29). Because 1,3-diaminopropane is not readily metabolized to the corresponding spermidine analog (30), the data suggest that putrescine has a negative effect independent of its conversion to spermidine. Both OrnDCase antizyme formation (20) and loss of OrnDCase protein (31) have been reported in cells treated with 1,3-diaminopropane.…”

Section: Discussionmentioning

confidence: 90%

Distinct roles of putrescine and spermidine in the regulation of ornithine decarboxylase in Neurospora crassa.

Davis¹,

Krasner²,

DiGangi³

et al. 1985

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

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We wished to identify metabolic signals governing changes in ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) activity in Neurospora crassa. By manipulations of the ornithine supply and by the use of inhibitors of the polyamine pathway, we found that spermidine negatively governs formation of active ornithine decarboxylase and that putrescine promotes inactivation of the enzyme. Direct addition of putrescine or spermidine to cycloheximide-treated cells confirmed the role of putrescine in enzyme inactivation and showed that spermidine had no effect on this process. Increases in ornithine decarboxylase activity caused by blocking spermidine synthesis occurred prior to a significant decrease in the spermidine pool. This is consistent with our prevous finding that only 10-20% of the spermidine pool is freely diffusible within N. crassa cells. We presume that only this small fraction of the pool is active in regulation.Ornithine decarboxylase (OrnDCase; L-ornithine carboxylyase, EC 4.1.1.17) is a key enzyme of the synthesis of polyamines (putrescine, spermidine, and spermine) in most fungi and animals (1-3). While the roles of polyamines in vivo are still uncertain, these compounds are indispensable in eukaryotes (2). An increase of OrnDCase activity and an increased rate of polyamine synthesis are correlated with the onset of rapid growth, of transformation to the neoplastic state, and of periods of cell differentiation in most organisms (see refs. in ref. 2). Moreover, OrnDCase is rapidly inactivated upon the cessation of growth or upon addition of polyamines to cells. The regulation of the amount of active enzyme is the major mechanism of control; feedback and product inhibition are weak or absent.A test of polyamine pathway intermediates ( Fig. 1) for their roles in regulating the OrnDCase of Neurospora crassa is reported here. We identify spermidine and putrescine as regulatory signals governing, respectively, the formation and inactivation of OrnDCase catalytic activity. The work is discussed in connection with our previous observations (4-6) that polyamines are highly compartmentalized in cells of N.crassa. MATERIALS AND METHODSStrains, Growth, and Sampling. The strain of N. crassa used was IC3, a prototrophic strain carrying the arginase-less (aga) allele, UM-906. Vogel's medium N (7) was used for growth. Cultures were grown exponentially in 1-liter aerated cultures (8) for determinations of growth rate, enzyme activity, and polyamine pools. Samples (20 ml) for dry weight were collected on Whatman no. 1 filter circles and dried with acetone. Mycelial samples (10 ml) for polyamine determinations were collected and washed on membrane filters (pore size, 5 ,um). The pad was extracted with 1 ml of 0.4 M perchloric acid with 1 mM EDTA. As an internal standard, 100 nmol of 1,7-diaminoheptane was added before centrifugation; the supernatant was saved and frozen until use. Mycelial samples (5 ml) for enzyme assay were permeabilized with a toluene/ethanol solution, followed by freezing (9).OrnD...

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Section: Discussionmentioning

confidence: 90%

Distinct roles of putrescine and spermidine in the regulation of ornithine decarboxylase in Neurospora crassa.

Davis¹,

Krasner²,

DiGangi³

et al. 1985

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

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“…The specificity of the enzyme is significantly different when compared to that of propylamine transferases from eubacteria [12] and eukaryotes [45]; indeed the enzyme from S . solfataricus is able to transfer the propylamine group from decarboxy-AdoMet to a primary amino group linked to a trimethylenic as well as to a tetramethylenic chain.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of propylamine transferase from Sulfolobus solfataricus, an extreme thermophilic archaebacterium

Cacciapuoti¹,

Porcelli²,

Cartenı́-Farina³

et al. 1986

Eur J Biochem

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The enzyme propylamine transferase, catalyzing the transfer of the propylamine moiety from S-adenosyl(5')-3-methylthiopropylamine to several amine acceptors, has been purified 643-fold in 20% yield from Sulfolobus solfaturicus, an extreme thermophilic archaebacterium optimally growing at 87 "C. The purified enzyme (specific activity 2.05 units/mg protein), is homogeneous by criteria of gel electrophoresis, gel filtration, isoelectric focusing and ultracentrifugation analysis. The molecular mass of the native enzyme was estimated to be about 110 kDa by gel permeation and ultracentrifugation analysis. The protein migrates on SDS/polyacrylamide gel electrophoresis as a single band of 35 kDa, suggesting that the enzyme is a trimer composed by identical subunits. An optimum pH of 7.5 and an acidic isoelectric point of 5.3 have been calculated. The optimum temperature was 90°C and no loss of activity is observable even after exposure of the purified enzyme to 100°C for 1 h. No reducing agents are required for enzymatic activity.Substrate specificity towards the amine acceptors is rather broad in that 1,3-diaminopropane ( K , = 1675 pM), putrescine (K, = 3850 pM), sym-norspermidine (K, = 954 pM) and spermidine (K, = 1539 pM) are recognized as substrates. Conversely S-adenosyl(5')-3-methylthiopropylamine is the only propylamine donor (K, = 7.9 pM) and the deamination of the sulfonium compound prevents the recognition by the enzyme. The reaction is irreversible and initial-rate kinetic studies indicate that the propylamine transfer is operated through a sequential mechanism. 5'-Methylthioadenosine, a product of the reaction, acts as a powerful competitive inhibitor with a Ki of 3.7 pM. Enzyme-substrate binding sites have been investigated with the aid of several substrate analogs and products. Among the compounds assayed, 5'-methylthiotubercidin, S-adenosyl(5')-3-thiopropylamine and S-adenosyl-3-thio-l,8-diaminooctane are the most active inhibitors.Propylamine transferases represent a group of enzymes catalyzing the transfer of the propylamine moiety from S-adenosyl-(5')-3-methylthiopropylamine (decarboxy-AdoMet) to appropriate amine acceptors [l -41. The reaction involves a nucleophilic attack on the methylene C-3 adjacent to the positively charged sulfur of decarboxy-AdoMet [5]. This class of enzymes is distributed in both prokaryotic and eukaryotic organisms. The recent development of rapid and sensitive methods for polyamine analysis [6 -81 and the utilization of affinity chromatography [9, 101 has made possible their purification and characterization : indeed spermidine Correspondence to V. Zappia, Istituto di Biochimica delle Macromolecole, I Facolta di Medicina e Chirurgia, Universita di Napoli, Via Costantinopoli 16, 1-80138, Napoli, ItalyAbbreviations. Decarboxy-AdoMet, S-adenosyl(S')-3-methylthiopropylamine; sym-norspermidine, 1,7-diamino-4-aza-eptane; symnorspermine, 1,l l-diamino-4,8-diaza-undecane; sym-homospermidine, 1,9-diamin0-5-aza-nonane; caldopentamine, 1,15-diamino- synthase has been purified from ra...

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“…1) was formed (71), and it was later shown that aminopropylcadaverine is as effective as Spd in restoring normal growth to a polyamine auxotroph (72). Production of aminopropylcadaverine by the AdoMetDC/SpdSyn pathway has also been demonstrated with mammalian and fungal cells (73,74).…”

Section: Polyamines In Bacteriamentioning

confidence: 99%

Polyamines in Eukaryotes, Bacteria, and Archaea

Michael

2016

Journal of Biological Chemistry

241

208

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Polyamines are primordial polycations found in most cells and perform different functions in different organisms. Although polyamines are mainly known for their essential roles in cell growth and proliferation, their functions range from a critical role in cellular translation in eukaryotes and archaea, to bacterial biofilm formation and specialized roles in natural product biosynthesis. At first glance, the diversity of polyamine structures in different organisms appears chaotic; however, biosynthetic flexibility and evolutionary and ecological processes largely explain this heterogeneity. In this review, I discuss the biosynthetic, evolutionary, and physiological processes that constrain or expand polyamine structural and functional diversity.The common feature of diverse polyamines found in eukaryotes, bacteria, and archaea is that they are all derived from amino acids and are positively charged at physiological pH. Structurally, they are mostly linear and flexible aliphatic chains containing two or more amine groups. They include the diamines 1,3-diaminopropane (Dap), 2 1,4-diaminobutane (putrescine, Put), and 1,5-diaminopentane (cadaverine, Cad), triamines sym-norspermidine (Nspd), spermidine (Spd), and sym-homospermidine (Hspd), the uncommon triamines aminopropylcadaverine and aminobutylcadaverine, the tetraamines norspermine (Nspm), spermine (Spm), and thermospermine (Tspm), and the uncommon tetraamine aminopropyl homospermidine (Fig. 1), and a wide range of longer chain polyamines and branched polyamines. This review will cover the distribution and biosynthesis of different polyamines in the three domains of life and will discuss the mechanisms underlying this biosynthetic diversity.

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Specificity of mammalian spermidine synthase and spermine synthase

Cited by 59 publications

References 33 publications

Distinct roles of putrescine and spermidine in the regulation of ornithine decarboxylase in Neurospora crassa.

Distinct roles of putrescine and spermidine in the regulation of ornithine decarboxylase in Neurospora crassa.

Purification and characterization of propylamine transferase from Sulfolobus solfataricus, an extreme thermophilic archaebacterium

Polyamines in Eukaryotes, Bacteria, and Archaea

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