Interaction of N-(DL-seryl)N′-(2,3,4-trihydroxybenzyl)-hydrazine with L-Dopa decarboxylase from pig kidney

Borri-Voltattorni, C; Minelli, Alba; Borri, P

doi:10.1007/bf02124039

Cited by 11 publications

(6 citation statements)

References 8 publications

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“…However, like many other pyridoxal phosphate enzymes, the enzyme also catalyses a "minor" reaction, referred to as decarboxylation-dependent transamination, which in fact was first observed by studying the effects of pig kidney DDC on ␣-methyl Dopa (1). A similar phenomena has been described for DDC when it acts on L-3,4-dihydroxyphenylalanine (L-Dopa) and m-tyrosine and their ␣-methyl derivatives (2, 3), D-5-hydroxytryptophan and D-tryptophan (4), and 5-hydroxytryptamine (5-HT, serotonin) (5); with all of the above substrates a progressive loss of decarboxylase activity is observed as a consequence of their interaction with the enzyme. The occurrence of abortive decarboxylation-transamination reactions is not uncommon among PLPdependent ␣-decarboxylases (6 -9).…”

Section: Pig Kidney Dopa Decarboxylase (Ddc) Expressed Insupporting

confidence: 62%

Mechanism-based Inactivation of Dopa Decarboxylase by Serotonin

Bertoldi

Moore

Maras³

et al. 1996

Journal of Biological Chemistry

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Pig kidney dopa decarboxylase (DDC) expressed in Escherichia coli is a homodimeric enzyme containing one catalytically active pyridoxal 5'-phosphate active site per subunit. In addition to catalyzing the decarboxylation of -aromatic amino acids, DDC also reacts with 5-hydroxytryptamine (5-HT), converting it to 5-hydroxyindolacetaldehyde and ammonia. These products have been identified by means of the enzymes alcohol dehydrogenase and glutamate dehydrogenase, together with high performance liquid chromatographic and mass spectroscopic analysis. The Kcat and Km values of this reaction were determined to be 0.48 min-1 and 0.47 mM, respectively. The NaBH4-reduced enzyme does not catalyze this reaction. Concurrent with this reaction, 5-HT inactivates DDC in both a time- and concentration-dependent manner and exhibits saturation of the rate of inactivation at high concentrations, with Ki and Kinact values of 0.40 mM and 0.023 min-1, respectively. Protection from inactivation by 5-HT was observed in the presence of the active site-directed inhibitor 3,4-dihydroxy-D-phenylalanine. Inactivation with [2-14C]5-HT results in the incorporation of 1 mol of label/enzyme subunit. Taken together, these findings indicate that 5-HT is both a substrate and a mechanism-based inactivator with a partition ratio for product formation versus inactivation of 21. The absorbance, CD, and fluorometric features of 5-HT-inactivated DDC have also been characterized. A speculative mechanism for the reaction and inactivation consistent with the experimental findings is presented.

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Section: Pig Kidney Dopa Decarboxylase (Ddc) Expressed Insupporting

confidence: 62%

Mechanism-based Inactivation of Dopa Decarboxylase by Serotonin

Bertoldi

Moore

Maras³

et al. 1996

Journal of Biological Chemistry

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“…The interaction of DDC with carbidopa, benserazide and Ro 4-5127 was investigated. While benserazide behaves as a poor inhibitor (K i ~ 0.3 mM) [158], either carbidopa [159] or Ro 4-5127 [160] bind to the enzyme by a hydrazone linkage with the PLP cofactor through its hydrazine moiety, and have been found to be powerful irreversible inhibitors. However, because hydrazine derivatives can react with both free PLP and all PLP-enzymes, these inhibitors are not selective for DDC.…”

Section: D-amino Acid Aminotransferasementioning

confidence: 99%

Pyridoxal 5-Phosphate Enzymes as Targets for Therapeutic Agents

Amadasi¹,

Bertoldi²,

Contestabile³

et al. 2007

CMC

177

157

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The vitamin B(6)-derived pyridoxal 5'-phosphate (PLP) is the cofactor of enzymes catalyzing a large variety of chemical reactions mainly involved in amino acid metabolism. These enzymes have been divided in five families and fold types on the basis of evolutionary relationships and protein structural organization. Almost 1.5% of all genes in prokaryotes code for PLP-dependent enzymes, whereas the percentage is substantially lower in eukaryotes. Although about 4% of enzyme-catalyzed reactions catalogued by the Enzyme Commission are PLP-dependent, only a few enzymes are targets of approved drugs and about twenty are recognised as potential targets for drugs or herbicides. PLP-dependent enzymes for which there are already commercially available drugs are DOPA decarboxylase (involved in the Parkinson disease), GABA aminotransferase (epilepsy), serine hydroxymethyltransferase (tumors and malaria), ornithine decarboxylase (African sleeping sickness and, potentially, tumors), alanine racemase (antibacterial agents), and human cytosolic branched-chain aminotransferase (pathological states associated to the GABA/glutamate equilibrium concentrations). Within each family or metabolic pathway, the enzymes for which drugs have been already approved for clinical use are discussed first, reporting the enzyme structure, the catalytic mechanism, the mechanism of enzyme inactivation or modulation by substrate-like or transition state-like drugs, and on-going research for increasing specificity and decreasing side-effects. Then, PLP-dependent enzymes that have been recently characterized and proposed as drug targets are reported. Finally, the relevance of recent genomic analysis of PLP-dependent enzymes for the selection of drug targets is discussed.

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“…It is interesting to note that this inactive form has properties quite similar to those found in pig kidney apoenzyme [3,21]. Further studies will be needed to elucidate the structural and functional basis of this absorbing form better.…”

Section: Pyridoxal-p Content and Its Absorption Spectral Propertiesmentioning

confidence: 98%

Purification and characterization of rat-liver 3,4-dihydroxyphenylalanine decarboxylase

Dominici

Tancini

Barra³

et al. 1987

Eur J Biochem

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A simple and rapid procedure, which takes advantage of the effectiveness of conventional and HPLC hydrophobic interaction, for the isolation of highly purified rat liver 3,4-dihydroxyphenylalanine decarboxylase is described in detail.Some of its structural and functional properties are reported and discussed in comparison with those of pig kidney 3,4-dihydroxyphenylalanine decarboxylase.Considerable efforts have been devoted during the recent years to characterize Dopa decarboxylase, a pyridoxal-P-dependent enzyme involved in the metabolism of aromatic amino acids. Most of these studies have been carried out on the enzyme purified from the kidney of either pig [l -31 or guinea pig [4], this tissue being an exceptionally rich source of Dopa decarboxylase.Dopa decarboxylase activity has also been detected in crude extracts of other peripheral organs, particularly liver and intestine [ S ] , but relatively little information is available about the enzyme from these sources, since no method yielding an homogeneous preparation has been published. Thus, we have developed a simple and rapid procedure, which takes advantage of the effectiveness of hydrophobic interaction chromatography, for the isolation of highly purified rat liver Dopa decarboxylase. This paper also reports some properties of this enzyme which are discussed in comparison with those of pig kidney Dopa decarboxylase. EXPERIMENTAL PROCEDURE MuteriulsThe following reagent grade chemicals were purchased from Sigma Chemical Co. : L-Dopa, L-Trp(S-OH), pyridoxal-P, 2,4,6-trinitrobenzene-l-sulfonic acid. DEAE-Bio-Gel was obtained from Bio-Rad; phenyl-Sepharose CL 4B was purchased from Pharmacia. All other chemicals were of the highest grade available.Hydrophobic interaction chromatography with HPLC was done on an LKB system gradient HPLC with a model 21 51 ultraviolet/visible detector, using a TSK phenyl-5 PW Correspondence to C. B. Voltattorni, Istituto Interfacolta di Chimica Biologica, Facolta di Farmacia e Scienze, Universita di Perugia, Via del Giochetto, 1-016100 Perugia, ItalyAbbreviations. Pyridoxal-P, pyridoxal 5'-phosphate; Dopa, 3,4-dihydroxyphenylalanine; Trp(5-OH), 5-hydroxytryptophan; mMe Dopa, a-methyl 3,4-dihydroxyphenylalanine.Enzyme. 3,4-Dihydroxyphenylalanine decarboxylase or aromatic L-amino acid decarboxylase (EC 4.1.1.28). column (internal diameter 7.5 x 75 mm) (Toyo Soda, Yamaguchi, Japan), a generous gift from LKB (Italy).Absorption spectra were recorded with a Cary 219 spectrophotometer. Enzyme assayEnzymatic activity was measured by the assay method of Sherald et al.[6], according to the modification introduced by Charteris and John [7]. The standard reaction mixture contained in a final volume of 250 pl, 100 p10.1 M potassium phosphate buffer, pH 6.8, 25 p1 10 pM pyridoxal-P and 25 pl 50 mM L-Dopa. The reaction, started by the addition of L-Dopa in order to reduce the non-enzymic reaction between pyridoxal-P and L-Dopa [l], was incubated 10 min at 25°C and then stopped by heating at 100°C for 1 min. Benzene (1.5 ml) and 2,4,6-trinit...

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Interaction of N-(DL-seryl)N′-(2,3,4-trihydroxybenzyl)-hydrazine with L-Dopa decarboxylase from pig kidney

Cited by 11 publications

References 8 publications

Mechanism-based Inactivation of Dopa Decarboxylase by Serotonin

Mechanism-based Inactivation of Dopa Decarboxylase by Serotonin

Pyridoxal 5-Phosphate Enzymes as Targets for Therapeutic Agents

Purification and characterization of rat-liver 3,4-dihydroxyphenylalanine decarboxylase

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