Assaying Ornithine and Arginine Decarboxylases in Some Plant Species

Birecka, H.; Bitonti, Alan J.; McCann, Peter P.

doi:10.1104/pp.79.2.509

Cited by 70 publications

(56 citation statements)

References 10 publications

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“…It is quite possible that pyrroline was produced but rapidly converted to GABA. Aminoguanidine, which inhibits Ptc breakdown in some plants (3,7) and animals (21), produced a small inhibition of Ptc breakdown in the pericarp tissue (Table III). It is likely that either AG did not fully get to the active site or the tomato diamine oxidase is somewhat different from that found in other plants.…”

Section: Gc-ms Analysismentioning

confidence: 99%

“…The conversion of Ptc into GABA suggests that a pathway involving a diamine oxidase may be operative in the tomato pericarp tissue: diamine oxidase produces y-aminobutyraldehyde (which spontaneously interconverts into pyrroline) which upon further oxidation results in GABA (7). This possibility was tested by application of AG, an inhibitor of diamine oxidase (3,7,21). Treatment with AG produced a small but significant inhibition of Ptc breakdown with a concomitant reduction in the formation of GABA, Glu, sugars, and organic acids (Table III).…”

Section: Gc-ms Analysismentioning

confidence: 99%

“…Samples (3)(4)(5) ,uL) were injected in split mode (50-1) onto a bonded methyl silicone capillary column (HP-1, 25 m x 0.2 mm).…”

Section: Gc-ms Analysismentioning

confidence: 99%

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Polyamine Metabolism in Ripening Tomato Fruit

Rastogi¹,

Davies²

1990

Plant Physiol.

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The metabolism of [1,4-14C]putrescine and [terminal methylene-3H]spermidine was studied in the fruit pericarp (breaker stage) discs of tomato (Lycopersicon esculentum Mill.) cv Rutgers, and the metabolites identified by high performance liquid chromatography and gas chromatography-mass spectrometry. The metabolism of both putrescine and spermidine was relatively slow; in 24 hours about 25% of each amine was metabolized.The 14C label from putrescine was incorporated into spermidine, 'y-aminobutyric acid (GABA), glutamic acid, and a polar fraction eluting with sugars and organic acids. In the presence of gabaculine, a specific inhibitor of GABA:pyruvate transaminase, the label going into glutamic acid, sugars and organic acids decreased by 80% while that in GABA increased about twofold, indicating that the transamination reaction is probably a major fate of GABA produced from putrescine in vivo.[3H]Spermidine was catabolized into putrescine and fl-alanine. The conversion of putrescine into GABA, and that of spermidine into putrescine, suggests the presence of polyamine oxidizing enzymes in tomato pericarp tissues. The possible pathways of putrescine and spermidine metabolism are discussed.The diamine Ptc2 and the PAs, Spd, and Spm are apparently ubiquitously distributed in plants, and changes in their levels and biosynthesis have been correlated with a variety of plant developmental processes (6,22,24).To determine the physiological role of PAs, it is important to understand how PA levels are regulated in plant tissues. Considerable attention has been given to PA biosynthesis, and the enzymes involved have been well-characterized in certain systems (6,22,24). PA breakdown, however, has been studied only in a few cases, and therefore relatively little is known of the metabolic fate of these compounds. It was previously reported that soybean seedlings labeled with [14C] Ptc produced GABA, Glu, Asp, and organic acids (15). Similarly, GABA, Glu, and Asp were also produced from Ptc by cultured radiata pine cotyledons (14). Terano and Suzuki (28,29) 2Abbreviations: Ptc, putrescine; PA(s), polyamine(s); Spd, spermidine; Spm, spermine; GABA, -y-aminobutyric acid; Dap, 1,3-diaminopropane; AG, aminoguanidine; GCU, gabaculine (3-amino 2, 3-dihydrobenzoic acid); MDL-7252 1, N'-methyl-N2-(buta-2,3-dienyl)butane-1 ,4-diamine.and Spm, and that of GABA from Spm, by corn shoots and seedlings, respectively. Also, pea shoots and oat leaves, respectively, were shown to oxidatively metabolize Ptc and Spd to GABA (7).We have been interested in investigating PA metabolism during tomato (Lycopersicon esculentum Mill.) fruit ripening, primarily because ethylene is involved in ripening (4) MATERIALS AND METHODS Plant MaterialThe seeds of tomato (Lycopersicon esculentum Mill.) cv Rutgers were germinated in 5 inch plastic pots in a mixture of peat and vermiculite (1:1 by weight), and seedlings with three to four leaves were transferred to 12 inch pots containing a mixture of prolite, vermiculite, and peat (1:1:2 by weight).Plants...

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Section: Gc-ms Analysismentioning

confidence: 99%

Section: Gc-ms Analysismentioning

confidence: 99%

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Polyamine Metabolism in Ripening Tomato Fruit

Rastogi¹,

Davies²

1990

Plant Physiol.

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“…The labeled Radiolabeled Chemicals, St. Louis, MO) was measured according to the method of Birecka et al (1985). The reaction mixtures contained 70 pL of enzymatic extract, 10 pL of labeled substrate (0.3 pCi), and unlabeled substrate, giving a final concentration of 5 mM of L-Arg or 10 mM of L-Orn, in a total volume of 150 pL.…”

Section: Extraction and Assay For Adc And O D C Activitymentioning

confidence: 99%

Arginase, Arginine Decarboxylase, Ornithine Decarboxylase, and Polyamines in Tomato Ovaries (Changes in Unpollinated Ovaries and Parthenocarpic Fruits Induced by Auxin or Gibberellin)

et al. 1996

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Arginase (EC 3.5.3.1) activity has been found in the ovaries and young fruits of tomato (Lycopersicon esculentum Mill. cv Rutgers). Changes in arginase, arginine decarboxylase (EC 4.1.1.1 9), and ornithine decarboxylase activity (EC 4.1 .I .17) and levels of free and conjugated putrescine, spermidine, and spermine were determined in unpollinated ovaries and in parthenocarpic fruits during the early stages of development induced by 2,4-dichlorophenoxyacetic acid (2,4-D) or gibberellic acid (CA,). Levels of arginase, free spermine, and conjugates of the three polyamines were constant in unpollinated ovaries and characteristic of a presenescent step. A marked decrease in arginase activity, free spermine, and polyamine conjugates was associated with the initiation of fruit growth due to cell division, and when cell expansion was initiated, the absence of arginase indicated a redirection of nitrogen metabolism to the synthesis of arginine. A transient increase in arginine decarboxylase and ornithine decarboxylase was also observed in 2,4-D-induced fruits. In general, 2,4-D treatments produced faster changes than CA,, and without treatment, unpollinated ovaries developed only slightly and senescence was hardly visible. Sensitivity to 2,4-D and CA, treatment remained for at least 2 weeks postanthesis.

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“…a-DL-Difluoromethylarginine, a, specific enzyme-activated inhibitor of arginine decarboxylase, at 0.1 to 2.0 millimolar had a weak inhibitory effect on the fungus. The effect was not dependent on the inhibitor concentration and there was no detectable arginine decarboxylase activity in the fungus.Our recent investigation on in vitro activities of ADC2 and ODC from higher plants (4,5) under the same conditions sporulation is three to four times as high as that found in cultures exposed to light (M. 0. Garraway, unpublished data).…”

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Inhibition of Ornithine Decarboxylase and Growth of the Fungus Helminthosporium maydis

Birecka¹,

Garraway²,

Baumann³

et al. 1986

Plant Physiol.

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a-DL-DifluoromethylomUithine (DFMO), a specific enzyme-activated inhibitor of ornithine decarboxylase, at 0.5 to 2.0 millimolar significantly inhibited mycelial growth and especially sporulation of Helminthosporium maydis in the dark; its inhibitory effect on sporulation was greatly increased under light conditions. Putrescine at 0.25 millimolar fully prevented the inhibitory effects of DFMO; the inhibition caused by the latter could not be prevented by cadaverine or CaC12. a-DL-Difluoromethylarginine, a, specific enzyme-activated inhibitor of arginine decarboxylase, at 0.1 to 2.0 millimolar had a weak inhibitory effect on the fungus. The effect was not dependent on the inhibitor concentration and there was no detectable arginine decarboxylase activity in the fungus.Our recent investigation on in vitro activities of ADC2 and ODC from higher plants (4,5) under the same conditions sporulation is three to four times as high as that found in cultures exposed to light (M. 0. Garraway, unpublished data). MATERIALS AND METHODSThe fungus was grown on a synthetic medium containing asparagine described previously (6). The sterilized agar culture media (20 ml per 100 x 20 mm sterile Petri dish) contained glucose, L-asparagine, KH2PO4, and MgSO4 at 0.5, 0.2, 0.3, and 0.15%. Sterilized solutions of Mn, Zn, Fe, and Cu salts at 1 ug/ 1 each were added at 0.2 ml per dish. Solutions of other compounds were filter sterilized prior to application. The pH of the culture media in experiment I was 5.0; in the remaining experiments it was adjusted to 6.0, which proved to be optimal for the fungal growth. Each Petri dish was inoculated at its center with a plug of mycelium 7 mm in diameter. Each experiment was carried out in four replicates. In experiments I and III the Petri dishes were placed in the dark at room temperature. In experiment II, 2 d after inoculation, the plates were transferred from darkness to diffused fluorescent light. Growth was determined by linear extension, and sporulation was expressed on dry weight basis (6).In experiment III the activities of ODC and ADC were determined after the enzymes were extracted with phosphate buffer (pH 7.6) (14) from mycelia exposed to the highest DFMO and DFMA concentrations. Repeated freezing and thawing, sonication, and maceration with glass beads was applied during extraction at 0 to 4°C. The enzymes were assayed as described previously (3,4)

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Assaying Ornithine and Arginine Decarboxylases in Some Plant Species

Cited by 70 publications

References 10 publications

Polyamine Metabolism in Ripening Tomato Fruit

Polyamine Metabolism in Ripening Tomato Fruit

Arginase, Arginine Decarboxylase, Ornithine Decarboxylase, and Polyamines in Tomato Ovaries (Changes in Unpollinated Ovaries and Parthenocarpic Fruits Induced by Auxin or Gibberellin)

Inhibition of Ornithine Decarboxylase and Growth of the Fungus Helminthosporium maydis

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